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Characterization of depth-related microbial communities in lake sediment by denaturing gradient gel electrophoresis of amplified 16S rRNA fragments 被引量:5
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作者 ZHAO Xingqing YANG Liuyan +4 位作者 YU Zhenyang PENG Naiying XIAO Lin YIN Daqiang QIN Boqiang 《Journal of Environmental Sciences》 SCIE EI CAS CSCD 2008年第2期224-230,共7页
The characterization of microbial communities of different depth sediment samples was examined by a culture-independent method and compared with physicochemical parameters, those are organic matter (OM), total nitro... The characterization of microbial communities of different depth sediment samples was examined by a culture-independent method and compared with physicochemical parameters, those are organic matter (OM), total nitrogen (TN), total phosphorus (TP), pH and redox potential (Eh). Total genomic DNA was extracted from samples derived from different depths. After they were amplified with the GC-341 f/907r primer sets of partial bacterial 16S rRNA genes, the products were separated by denaturing gradient gel electrophoresis (DGGE). The profile of DGGE fingerprints of different depth sediment samples revealed that the community structure remained relatively stable along the entire 45 cm sediment core, however, principal-component analysis of DGGE patterns revealed that at greater sediment depths, successional shifts in community structure were evident. The principle coordinates analysis suggested that the bacterial communities along the sediment core could be separated into two groups, which were located 0-20 cm and 21-45 cm, respectively. The sequencing dominant bands demonstrated that the major phylogenetic groups identified by DGGE belonged to Bacillus, Bacterium, Brevibacillus, Exiguobacterium, γ-Proteobacterium, Acinetobacter sp. and some uncultured or unidentified bacteria. The results indicated the existence of highly diverse bacterial community in the lake sediment core. 展开更多
关键词 16S rRNA denaturing gradient gel electrophoresis (DGGE) Lake Taihu microbial diversity SEDIMENT vertical distribution
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Analysis of interspecies adherence of oral bacteria using a membrane binding assay coupled with polymerase chain reaction-denaturing gradient gel electrophoresis profiling 被引量:1
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作者 Ren-ke Wang Xue-song He +4 位作者 Wei Hu Renate Lux Ji-yao Li Xue-dong Zhou Wen-yuan Shi 《International Journal of Oral Science》 SCIE CAS CSCD 2011年第2期90-97,共8页
Information on co-adherence of different oral bacterial species is important for understanding interspecies interactions within oral microbial community. Current knowledge on this topic is heavily based on pariwise co... Information on co-adherence of different oral bacterial species is important for understanding interspecies interactions within oral microbial community. Current knowledge on this topic is heavily based on pariwise coaggregation of known, cultivable species. In this study, we employed a membrane binding assay coupled with polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) to systematically analyze the co-adherence profiles of oral bacterial species, and achieved a more profound knowledge beyond pairwise coaggregation. Two oral bacterial species were selected to serve as "bait": Fusobacterium nucleatum (F. nucleatum) whose ability to adhere to a multitude of oral bacterial species has been extensively studied for pairwise interactions and Streptococcus mutans (S. mutans) whose interacting partners are largely unknown. To enable screening of interacting partner species within bacterial mixtures, cells of the "bait" oral bacterium were immobilized on nitrocellulose membranes which were washed and blocked to prevent unspecific binding. The "prey" bacterial mixtures (including known species or natural saliva samples) were added, unbound ceils were washed off after the incubation period and the remaining cells were eluted using 0.2 mol.L1 glycine. Genomic DNA was extraeted, subjeeted to 16S rRNA PCR amplification and separation of the resulting PCR produets by DGGE. Selected bands were recovered from the gel, sequenced and identified via Nucleotide BLAST searches against different databases. While few bacterial species bound to S. mutans, consistent with previous findings F.. nucleatum adhered to a variety of bacterial species including uncultivable and uneharacterized onesl This new approach can more effectively analyze the co-adherence profiles of oral bacteria, and could facilitate the systematic study of interbacterial binding of oral microbial species. 展开更多
关键词 membrane binding assay polymerase chain reaction-denaturing gradient gel electrophoresis COAGGREGATION Fusobacterium nucleatum Streptococcus mutans
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Denaturing gradient gel electrophoresis fingerprinting of soil bacteria in the vicinity of the Chinese Great Wall Station, King George Island, Antarctica
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作者 Qi Pan Feng Wang +3 位作者 Yang Zhang Minghong Cai Jianfeng He Haizhen Yang 《Journal of Environmental Sciences》 SCIE EI CAS CSCD 2013年第8期1649-1655,共7页
Bacterial diversity was investigated in soil samples collected from 13 sites around the Great Wall Station, Fildes Peninsula, King George Island, Antarctica, using denaturing gradient gel electrophoresis (DGGE) of 1... Bacterial diversity was investigated in soil samples collected from 13 sites around the Great Wall Station, Fildes Peninsula, King George Island, Antarctica, using denaturing gradient gel electrophoresis (DGGE) of 16S rRNA genes. The classes α-, β-, and γ-Proteobacteria, as well as the phylum Actinobacteria, were found to be the dominant bacteria in the soils around the Great Wall Station. Although the selected samples were not contaminated by oil, a relationship between soil parameters, microbial biodiversity, and human impact was still seen. Sample sites in human impacted areas showed lower bacterial biodiversity (average H′= 2.65) when compared to nonimpacted sites (average H′= 3.05). There was no statistically significant correlation between soil bacterial diversity and total organic carbon (TOC), total nitrogen, or total phosphorus contents of the soil. Canonical correlation analysis showed that TOC content was the most important factor determining bacterial community profiles among the measured soil parameters. In conclusion, microbial biodiversity and community characteristics within relatively small scales (1.5 km) were determined as a function of local environment parameters and anthropogenic impact. 展开更多
关键词 ANTARCTICA 16S rRNA gene denaturing gradient gel electrophoresis human impact
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Variations in Laboratory-Scale Actinomycete Communities Exposed to Cadmium as Assessed by Denaturing Gradient Gel Electrophoresis Profiles
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作者 YUAN Hai-Ping,MIN Hang 2,LIU Ji,YAN Bo and L Zhen-Mei College of Life Science,Zhejiang University,Hangzhou 310058 (China) 《Pedosphere》 SCIE CAS CSCD 2010年第2期174-184,共11页
The actinomycete populations and functions in cadmium (Cd) contaminated soil were investigated by the cultivation- independent molecular methods. The genomic DNA was extracted and purified from soil adulterated with... The actinomycete populations and functions in cadmium (Cd) contaminated soil were investigated by the cultivation- independent molecular methods. The genomic DNA was extracted and purified from soil adulterated with various con- centrations of Cd in the laboratory. The partial 16S rDNA genes were amplified by polymerase chain reaction (PCR) using specific primers bound to evolutionarily conserved regions within these actinomycete genes. The diversity in PCR- amplified products, as measured by denaturing gradient gel electrophoresis (EGGE), was used as a genetic fingerprint of the population. Principle component analysis and Shannon-Weaver diversity index (H) analyses were used to analyze the DGGE results. Results showed that the two principal components accounted for only a low level of the total variance. The value H in contaminated soil was lower than that in the control at later stages of cultivation, whereas at earlier stages it was higher. Among the six sampling time points, the first, fifth and sixth weeks had the highest values of H. Significantly negative correlations between bioavallable Cd concentration and H values existed in the samples from weeks 2 (R = 0.929, P 〈 0.05) and 4 (R = 0.909, P 〈 0.05). These results may shed light on the effect of Cd on the soil environment and the chemical behavior and toxicity of Cd to actinomycetes. 展开更多
关键词 denaturing gradient gel electrophoresis GENES principal component analysis Shannon-Weaver diversity index
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Monitoring of microbial community structure and succession in the biohydrogen production reactor by denaturing gradient gel electrophoresis(DGGE) 被引量:5
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作者 XING Defeng REN Nanqi +2 位作者 GONG Manli LI Jianzheng LI Qiubo 《Science China(Life Sciences)》 SCIE CAS 2005年第2期155-162,共8页
To study the structure of microbial communities in the biological hydrogen produc-tion reactor and determine the ecological function of hydrogen producing bacteria,anaerobic sludge was obtained from the continuous sti... To study the structure of microbial communities in the biological hydrogen produc-tion reactor and determine the ecological function of hydrogen producing bacteria,anaerobic sludge was obtained from the continuous stirred tank reactor(CSTR)in different periods of time,and the diversity and dynamics of microbial communities were investigated by denaturing gra-dient gel electrophoresis(DGGE).The results of DGGE demonstrated that an obvious shift of microbial population happened from the beginning of star-up to the 28th day,and the ethanol type fermentation was established.After 28 days the structure of microbial community became stable,and the climax community was formed.Comparative analysis of 16S rDNA sequences from reamplifying and sequencing the prominent bands indicated that the dominant population belonged to low G+C Gram-positive bacteria(Clostridium sp.and Ethanologenbacterium sp.),β-proteobacteria(Acidovorax sp.),γ-proteobacteria(Kluyvera sp.),Bacteroides(uncultured bacte-rium SJA-168),and Spirochaetes(uncultured eubacterium E1-K13),respectively.The hydrogen production rate increased obviously with the increase of Ethanologenbacterium sp.,Clostridium sp.and uncultured Spirochaetes after 21 days,meanwhile the succession of ethanol type fer-mentation was formed.Throughout the succession the microbial diversity increased however it decreased after 21 days.Some types of Clostridium sp.Acidovorax sp.,Kluyvera sp.,and Bac-teroides were dominant populations during all periods of time.These special populations were essential for the construction of climax community.Hydrogen production efficiency was de-pendent on both hydrogen producing bacteria and other populations.It implied that the co-metabolism of microbial community played a great role of biohydrogen production in the reactors. 展开更多
关键词 biohydrogen production microbial communities 16S rRNA denaturing gradient gel electrophoresis(DGGE).
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Compressive Mechanical Characteristics of Multi-layered Gradient Hydroxyapatite Reinforced Poly(Vinyl Alcohol) Gel Biomaterial
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作者 Yusong Pan Qianqian Shen +1 位作者 Chengling Pan Jing Wang 《Journal of Materials Science & Technology》 SCIE EI CAS CSCD 2013年第6期551-556,共6页
Functional gradient materials provided us a new concept for artificial articular cartilage design with gradient component and gradient structure where one side of the material is high free water content thereby provid... Functional gradient materials provided us a new concept for artificial articular cartilage design with gradient component and gradient structure where one side of the material is high free water content thereby providing excellent lubrication function and the opposite side of the material is high hydroxyapatite content, thereby improving the bioactivity of the material and stimulating cell growth. The goal of the present study was to develop a multi-layered gradient HA/PVA gel biocomposites through layer-by-layer casting method combing with freeze/thaw cycle technology. The various influence factors on the compressive strength and modulus of the multi-layered gradient biocomposites were investigated. The results showed that the compressive mechanical characteristics of the biocomposites were similar to that of natural articular cartilage. Both the compressive strength and modulus of the multi-layered gradient HA/PVA gel biocomposites increased exponentially with the rise of compressive strain ratio. Both the compressive strength and average compressive modulus of the biocomposites improved with the rise of freeze/thaw cycle times and total concentration of HA particles in the biocomposites, but they showed decreasing tendency with the rise of HA concentration difference between adjacent layers. 展开更多
关键词 Multi-layered gradient HA/PVA gel biocomposites Freeze/thaw cycle times Strength MODULUS
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Succession of Denitrifying Community Composition in Coastal Wetland Soils Along a Salinity Gradient 被引量:8
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作者 PIAO Zhe ZHANG Wen-Wen +2 位作者 MA Shuai LI Yu-Min YIN Shi-Xue 《Pedosphere》 SCIE CAS CSCD 2012年第3期367-374,共8页
To better understand the effect of salinity on denitrification communities, soils along a salinity gradient (ranging from 7.32 to 1.70 mS cm 1) in a wetland along the Yellow Sea coastline in Jiangsu Province, China,... To better understand the effect of salinity on denitrification communities, soils along a salinity gradient (ranging from 7.32 to 1.70 mS cm 1) in a wetland along the Yellow Sea coastline in Jiangsu Province, China, were studied using both culture-dependent and -independent methods. Culture efforts yielded 82 isolates in total, 81.7% of which were close relatives of Bacillus sp. based on partial sequences of their 16S rRNA genes. Denaturing gradient gel electrophoresis (DGGE) analysis based on 16S rRNA sequences suggested possible existence of bacterial community succession along the salinity gradient. Clone library analysis based on nosZ gene sequences (coding nitrous oxide reductase) showed that operational taxonomic units (OTUs) associated with α-proteobacteria dominated in all three soils, whereas those associated with β- and γ-subdivisions showed a clear succession. In the high salinity soil, only the OTUs associated with a-subdivision were found. In the medium salinity soil, small proportions of β- (6.5%) and γ-associated (19.6%) OTUs were found. In the low salinity soil, the proportions were further increased to 33% and 25% for β- and γ-Proteobacteria, respectively. Statistic analysis using Unifrac P test showed that nosZ-communities in different saline soils were significantly different from each other. It could be concluded that α-subdivision of nosZ-community tended to be sustained in high salinity environments whereas β and γ-subdivisions, especially the former, tended to be sustained in low salinity environments. Salinity was the key determinant of nosZ-community composition in the environment. 展开更多
关键词 clone library denaturing gradient gel electrophoresis (DGGE) nosZ gene Yellow Sea coastline
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Preparation and dielectric properties of compositionally graded (Ba,Sr)TiO_3 thin film by sol-gel technique 被引量:2
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作者 章天金 王军 +3 位作者 张柏顺 王今朝 万能 胡兰 《中国有色金属学会会刊:英文版》 CSCD 2006年第B01期119-122,共4页
Compositional graded BaxSr1-xTiO3 (x=0.6, 0.7, 0.8, 0.9, 1.0) (BST) thin films (less than 400 nm) were fabricated on Si and Pt/Ti/SiO2/Si substrates by sol-gel technique. A special heating treatment was employed to fo... Compositional graded BaxSr1-xTiO3 (x=0.6, 0.7, 0.8, 0.9, 1.0) (BST) thin films (less than 400 nm) were fabricated on Si and Pt/Ti/SiO2/Si substrates by sol-gel technique. A special heating treatment was employed to form the uniform composition gradients at 700 ℃. The microstructures of the films were studied by means of X-ray diffraction, atomic force microscope and field emission scanning electron microscopy. The results show that the films have uniform and crack-free surface morphology with perovskite structure phase. The small signal dielectric constant (εr) and dielectric loss (tan δ) are found to be 335 and 0.045 at room temperature and 200 kHz. The dielectric properties change significantly with applied dc bias, and the graded thin film show high tunability of 42.3% at an applied field of 250 kV/cm. All the results indicate that the graded BST thin films prepared by sol-gel technique have a promising candidate for microelectronic device. 展开更多
关键词 (Ba Sr)TiO3薄膜 基质 溶胶-凝胶法 制备 介电性质 成分梯度
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高通量测序和DGGE分析土壤微生物群落的技术评价 被引量:163
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作者 夏围围 贾仲君 《微生物学报》 CAS CSCD 北大核心 2014年第12期1489-1499,共11页
【目的】比较新一代高通量测序与传统的变性梯度凝胶电泳(Denaturing Gradient Gel Electrophoresis,DGGE)指纹图谱技术,评价两种技术研究土壤微生物群落结构的优缺点。【方法】针对新西兰典型草地和森林土壤,以16S rRNA基因为标靶,通... 【目的】比较新一代高通量测序与传统的变性梯度凝胶电泳(Denaturing Gradient Gel Electrophoresis,DGGE)指纹图谱技术,评价两种技术研究土壤微生物群落结构的优缺点。【方法】针对新西兰典型草地和森林土壤,以16S rRNA基因为标靶,通过高通量测序和DGGE技术分析土壤微生物群落的组成、丰度和多样性,比较两种方法在土壤微生物研究中的适用性。【结果】在不同的微生物分类水平,高通量测序草地土壤检测到22门,54纲,60目,131科,350属;而DGGE仅检测到6门,9纲,8目,10科,10属,表明DGGE显著低估了土壤微生物的群落组成。森林土壤也得到了类似规律,高通量测序的检测灵敏度是DGGE的3.8、6.7、6.4、19.2及39.4倍。进一步分析土壤中主要微生物类群的相对丰度,发现分类水平越低,高通量测序与DGGE的结果差异越大,尤其在科和属的水平上差异最大。以高通量测序结果为标准,DGGE明显高估了土壤中大多数微生物类群的相对丰度,最高可达2000倍。两种方法都表明草地土壤的多样性指数高于森林土壤,但DGGE多样性指数的绝对值远低于高通量测序结果。【结论】高通量测序能够较为全面和准确的反映土壤微生物群落结构,而DGGE仅能够反映有限的优势微生物类群,在很大程度上极可能低估土壤微生物的物种组成并高估其丰度。 展开更多
关键词 高通量测序 变性梯度凝胶电泳(Denaturing gradient gel Electrophoresis DGGE) 土壤微生物群落
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赣南稀土矿山废弃地的氨氮污染和微生物多样性分析
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作者 洪跃辉 冯爱娟 王江海 《广东化工》 2025年第7期106-108,124,共4页
目的:揭示赣南稀土矿山废弃地的氨氮污染情况和微生物多样性。方法:在稀土矿山废弃地采集16个土壤样品,测定氨氮含量;提取基因组DNA后,采用PCR扩增16SrRNA基因和18SrRNA基因片段,然后进行变性梯度凝胶电泳(DGGE)分析。结果:土壤的氨氮... 目的:揭示赣南稀土矿山废弃地的氨氮污染情况和微生物多样性。方法:在稀土矿山废弃地采集16个土壤样品,测定氨氮含量;提取基因组DNA后,采用PCR扩增16SrRNA基因和18SrRNA基因片段,然后进行变性梯度凝胶电泳(DGGE)分析。结果:土壤的氨氮平均浓度介于483.2 mg/kg~899.4 mg/kg,样品的细菌种群Shannon-Weaver系数(H)介于0.12~2.39,物种丰度系数(S)介于0.10~1.33。真菌种群H值介于1.09~2.59,S值介于0.26~1.46。细菌和真菌的H值和S值均偏小,DGGE电泳条带不多。结论:采样区存在严重的氨氮污染,在高浓度氨氮的影响下,微生物丰度低,群落结构简单,缺乏多样性,真菌多样性比细菌稍好,真菌群落多样性遭受破坏的程度比细菌小。 展开更多
关键词 赣南 变性梯度凝胶电泳 微生物多样性 稀土矿山废弃地
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PCR-DGGE Analysis of Bacterial Communities Structure in Babylonia areolata Culture Systems of The Subtidal Zone and The Pond Mulched Plastic Film and Sand in Bottom
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作者 李淑芳 邱德全 +2 位作者 张继东 杨世平 邱明生 《Agricultural Science & Technology》 CAS 2015年第7期1535-1539,1543,共6页
To know the bacterial communities structure in Babylonia areolata culture systems and to research and optimize the management pattem of Babylonia areola-ta culture systems of the pond mulched plastic film and sand in ... To know the bacterial communities structure in Babylonia areolata culture systems and to research and optimize the management pattem of Babylonia areola-ta culture systems of the pond mulched plastic film and sand in bottom, the bacte- rial communities in Babylonia areolata culture systems of the sub-tidal zone and the pond mulched plastic film and sand in bottom were analyzed at molecular level by adopting the denaturing gradient gel electrophoresis (DGGE). The results indicated that the dominant bacterial communities in Babylonia areolata culture systems of the sub-tidal zone and the pond mulched plastic film and sand in bottom, which were built on the basis of the seawater in East-island of Zhanjiang, included Proteobac- teda Chloroflexi, Cyanobacteria and Actinobacteria. The dominant bacterial groups in the above pond culture system were Garnmaproteobacteria, Alphaproteobacteria, Deltaprotecbacteda, Epsilonproteobacteda, Anaerolineae, Cyanobacteria and Acti- nobacteda. The dominant bacterial communities in the subtidal zone culture system were Gammaprotecbacteda, Alphaproteobacteria, Deltaproteobacteria, Anaerolineae and Cyanobacteda, and there were less Epsilonproteobacteria and Actinobacteria in the culture system. The higher diversity was detected in the above two culture sys- tems. The results of unweighted pair group method with arithmetic average (UPG- MA) showed that the bacterial communities of the sediment samples S1 and S2 in the above two culture systems were a cluster, the similarity of bacterial communities was 54.5%. The bacterial communities of seawater samples S3 and S4 in the above culture systems were in clusters, and the similarity of the bacterial communi- ties was 84.0%. The results showed that the microorganism ecological level in the Babylonia areolata culture systems of the pond mulched plastic film and sand in bottom could be similar to the sub-tidal zone culture systems through changing the pond seawater and monitoring the microbial population. 展开更多
关键词 Bacterial communities structure Denaturing gradient gel electrophoresis (DGGE) Culture system of the sub-tidal zone Culture system of the pond mulched plastic film and sand Babylonia areolata
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PCR-DGGE技术用于湖泊沉积物中微生物群落结构多样性研究 被引量:80
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作者 赵兴青 杨柳燕 +6 位作者 陈灿 肖琳 蒋丽娟 马喆 朱昊巍 于振洋 尹大强 《生态学报》 CAS CSCD 北大核心 2006年第11期3610-3616,共7页
采用PCR-DGGE分子指纹图谱技术比较南京市玄武湖、奠愁湖和太湖不同位置的表层沉积物微生物群落结构,研究结果表明,三湖泊沉积物微生物的16SrDNA的PCR扩增结果约为626bp,为16S rDNA V3~V5区特异性片段。玄武湖和莫愁湖表层沉积物中... 采用PCR-DGGE分子指纹图谱技术比较南京市玄武湖、奠愁湖和太湖不同位置的表层沉积物微生物群落结构,研究结果表明,三湖泊沉积物微生物的16SrDNA的PCR扩增结果约为626bp,为16S rDNA V3~V5区特异性片段。玄武湖和莫愁湖表层沉积物中大约有20种优势菌群,且同一湖泊不同采样点DGGE图谱的差异性不大,细菌群落结构具有较高的相似性,而太湖样品DGGE条带的数目和位置表现出明显差异,且不同采样点图谱的差异性较大。三湖泊除具有特征性的微生物种属外,还分布约5个相同的细菌种群,可能与沉积物的理化性质和水生植被的影响相关。对DGGE图谱中7条主带进行回收、扩增和测序,结果显示其优势菌群具有不同的序列组成,其中5个序列与Genebank中已登录的细菌种群的同源性≥99%,2个序列的同源性为96%和93%,其中2个相似的细菌类群目前尚未获得纯培养。 展开更多
关键词 沉积物 微生物多样性 变性梯度凝胶电泳(DGGE) 16S RDNA 序列测定
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PCR-DGGE技术在农田土壤微生物多样性研究中的应用 被引量:84
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作者 罗海峰 齐鸿雁 +1 位作者 薛凯 张洪勋 《生态学报》 CAS CSCD 北大核心 2003年第8期1570-1575,共6页
变性梯度凝胶电泳技术 ( DGGE)在微生物生态学领域有着广泛的应用。研究采用化学裂解法直接提取出不同农田土壤微生物基因组 DNA,并以此基因组 DNA为模板 ,选择特异性引物 F357GC和 R518对 1 6Sr RNA基因的 V3区进行扩增 ,长约 2 30 bp... 变性梯度凝胶电泳技术 ( DGGE)在微生物生态学领域有着广泛的应用。研究采用化学裂解法直接提取出不同农田土壤微生物基因组 DNA,并以此基因组 DNA为模板 ,选择特异性引物 F357GC和 R518对 1 6Sr RNA基因的 V3区进行扩增 ,长约 2 30 bp的 PCR产物经变性梯度凝胶电泳 ( DGGE)进行分离后 ,得到不同数目且分离效果较好的电泳条带。结果说明 ,DGGE能够对土壤样品中的不同微生物的 1 6Sr RNA基因的 V3区的 DNA扩增片断进行分离 ,为这些 DNA片断的定性和鉴定提供了条件。与传统的平板培养方法相比 ,变性梯度凝胶电泳 ( DGGE)技术能够更精确的反映出土壤微生物多样性 ,它是一种有效的微生物多样性研究技术。 展开更多
关键词 变性梯度凝胶电泳 DGGE 基因组DNA 16S RRNA 微生物多样性 农田土壤
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秸秆还田后土壤微生物群落结构变化的初步研究 被引量:38
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作者 赵勇 李武 +3 位作者 周志华 张晓君 潘迎捷 赵立平 《农业环境科学学报》 CAS CSCD 北大核心 2005年第6期1114-1118,共5页
通过实验室条件下的小麦秸秆粉和油菜秸秆粉的还土试验,结合常规的分析手段和基于DNA的分子技术(变性梯度凝胶电泳(DGGE)及测序技术),初步研究了秸秆粉还田后土壤物理化学特性及生物学特性的变化。结果表明,培养60d后,秸秆还田土壤的肥... 通过实验室条件下的小麦秸秆粉和油菜秸秆粉的还土试验,结合常规的分析手段和基于DNA的分子技术(变性梯度凝胶电泳(DGGE)及测序技术),初步研究了秸秆粉还田后土壤物理化学特性及生物学特性的变化。结果表明,培养60d后,秸秆还田土壤的肥力明显提高,其纤维素酶活性明显增强。DGGE图谱表明,对照土壤(S)以及处理土壤(SW和SR),在培养过程中,β-Proteobacteria类细菌组成都在发生变化。对其中一个样品的DGGE条带进行了割胶测序,测序结果表明,大部分条带代表的微生物是未培养的或不可培养的。采用传统分离培养技术,从秸秆还田土壤中分离了2株纤维素降解菌。以上结果说明,秸秆还田具有良好的土壤综合效应。 展开更多
关键词 秸秆 纤维素酶活性 变性梯度凝胶电泳 β-Proteobacteria 纤维素降解菌
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复合垂直流人工湿地系统中不同植物根际微生物群落结构 被引量:48
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作者 雷旭 李冰 +2 位作者 李晓 王璐 朱健 《生态学杂志》 CAS CSCD 北大核心 2015年第5期1373-1381,共9页
为了研究复合垂直流人工湿地在夏季和秋季运行期间湿地中不同植物根际微生物群落的结构特征,采用PCR-DGGE技术和基因测序技术,对复合垂直流人工湿地系统中梭鱼草(Pontederia cordata)、美人蕉(Canna indica)、再力花(Thalia dealbata)3... 为了研究复合垂直流人工湿地在夏季和秋季运行期间湿地中不同植物根际微生物群落的结构特征,采用PCR-DGGE技术和基因测序技术,对复合垂直流人工湿地系统中梭鱼草(Pontederia cordata)、美人蕉(Canna indica)、再力花(Thalia dealbata)3种植物根际微生物群落的结构特征和组成多样性进行分析。结果表明,3种植物根际微生物的种类和数量各不相同。人工湿地下行池中梭鱼草、美人蕉、再力花根际微生物的Shannon指数分别为2.91、2.96和2.90,说明美人蕉根际能更好地富集周围的微生物,提高微生物群落的多样性。通过比较各种植物根际微生物的相似度,发现根际微生物群落结构存在一定的季节差异;美人蕉和再力花根际微生物群落结构相似度较高(65%以上)。微生物群落组成多样性的结果显示,复合垂直流人工湿地系统中3种植物在实验期间根际微生物群落分属8个纲:α-变形菌纲(14.37%)、β-变形菌纲(38.13%)、γ-变形菌纲(6.87%)、ε-变形菌纲(16.87%)、δ-变形菌纲(6.87%)、放线菌纲(8.76%)、拟杆菌纲(4.38%)、硝化螺旋菌纲(3.75%)。β-变形菌纲为该人工湿地系统试验期间不同植物根际富集的主要细菌类群。本实验为以后人工湿地系统植物优化选择提供了一定的理论依据。 展开更多
关键词 人工湿地 植物根际 DGGE技术 微生物群落
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电针对溃疡性结肠炎大鼠肠道微生态的保护作用 被引量:110
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作者 侯天舒 韩晓霞 +4 位作者 杨阳 赵纪岚 任亚东 余曙光 吴巧凤 《针刺研究》 CAS CSCD 北大核心 2014年第1期27-34,共8页
目的:观察电针对溃疡性结肠炎(UC)大鼠肠道菌群的影响,从肠道微生态的角度探讨电针治疗UC的机制。方法:39只健康雄性SD大鼠随机分为空白组(13只)和UC造模组(26只),造模组饮用4%的葡聚糖硫酸钠(DSS)溶液7d复制UC模型后随机分为模型组和... 目的:观察电针对溃疡性结肠炎(UC)大鼠肠道菌群的影响,从肠道微生态的角度探讨电针治疗UC的机制。方法:39只健康雄性SD大鼠随机分为空白组(13只)和UC造模组(26只),造模组饮用4%的葡聚糖硫酸钠(DSS)溶液7d复制UC模型后随机分为模型组和电针组。电针"天枢""足三里""上巨虚"穴,每日1次,每次10min,治疗5d。观察实验过程中大鼠疾病活动指数的变化;收集各组大鼠粪便排泄物,用多聚酶链反应-变性梯度凝胶电泳法分析肠道菌群的多样性、相似性及丰度值,对优势条带进行回收测序,采用Quantity one等软件对所得结果图谱进行分析,对优势条带进行菌种鉴定及含量分析。结果:模型大鼠肠道菌群的丰度值及多样性指数明显降低,疾病活动指数明显增高(P<0.05,P<0.01)。电针能降低UC模型大鼠疾病活动指数,提高肠道菌群的丰度值及多样性指数(P<0.05,P<0.01);菌种鉴定表明电针能提高UC模型大鼠肠道菌群中乳酸杆菌(Lactobacillus sp.)和毛螺科菌(Lachnospiraceae bacterium)含量,降低双酶梭菌(Clostridium bifermentans)的含量(P<0.05)。结论:电针能明显改善UC模型大鼠肠道菌群的多样性以及有益菌群的含量,对UC模型大鼠肠道微生态发挥的保护作用很可能是电针治疗UC的机制之一。 展开更多
关键词 溃疡性结肠炎 电针干预 肠道微生态 多聚酶链反应-变性梯度凝胶电泳
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间作大豆对甘蔗根际土壤细菌及固氮菌多样性的影响 被引量:33
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作者 彭东海 杨建波 +4 位作者 李健 邢永秀 覃刘东 杨丽涛 李杨瑞 《植物生态学报》 CAS CSCD 北大核心 2014年第9期959-969,共11页
为探讨间作大豆(Glycine max)对甘蔗(Saccharum officinarum)根际土壤细菌及固氮细菌多样性的影响,收集和开发固氮菌资源,筛选高效甘蔗联合固氮体系,选用3个甘蔗栽培品种‘ROC22’、‘GT21’、‘B8’与大豆品种‘Guizao 2’进行间种栽培... 为探讨间作大豆(Glycine max)对甘蔗(Saccharum officinarum)根际土壤细菌及固氮细菌多样性的影响,收集和开发固氮菌资源,筛选高效甘蔗联合固氮体系,选用3个甘蔗栽培品种‘ROC22’、‘GT21’、‘B8’与大豆品种‘Guizao 2’进行间种栽培,采用巢式PCR特异扩增细菌16S rRNA基因片段和固氮细菌nifH基因片段,并结合变性梯度凝胶电泳(DGGE)技术,对间作大豆的甘蔗根际土壤细菌及固氮细菌进行系统演化和多样性分析。聚类分析结果显示,间作大豆改变了甘蔗根际土壤细菌及固氮细菌原来的群落组成结构,尤其对固氮菌群落组成的改变更大,但对群落物种的优势度影响较小。Shannon-Wiener多样性指数和Simpson多样性指数分析结果表明,甘蔗-大豆间作显著影响甘蔗根际土壤中细菌和固氮菌的多样性,其中对固氮细菌多样性的影响较大。不同甘蔗品种的根际土壤细菌和固氮菌在间作大豆条件下表现出不同的多样性,‘ROC22’和‘GT21’间作处理甘蔗根际土壤固氮细菌的Shannon-Wiener多样性指数显著高于单作处理,而‘ROC22’与大豆间作处理的甘蔗根际土壤固氮菌多样性最为丰富。在大豆生长盛期,间作处理的甘蔗根际土壤细菌多样性最为丰富,不同处理间的差异也最大,随后下降。总体来看,甘蔗-大豆间作显著地影响根际土壤细菌和固氮菌的群落结构和群落多样性,有助于对甘蔗合理间作栽培模式的认识和筛选高效甘蔗联合固氮体系。 展开更多
关键词 变性梯度凝胶电泳 间作 固氮细菌 大豆 甘蔗
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大港孔店油田水驱油藏微生物群落的分子分析 被引量:37
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作者 佘跃惠 张学礼 +2 位作者 张凡 王凌华 赵立平 《微生物学报》 CAS CSCD 北大核心 2005年第3期329-334,共6页
通过多聚酶链式反应 温度梯度凝胶电泳(PCR TGGE)和构建16SrRNA基因克隆文库两种方法对比研究了大港油田孔二北断块注水井和采油井的微生物群落结构。16SrDNAV3区PCR扩增产物的TGGE图谱分析表明,这两个油井的微生物群落结构差异很大。... 通过多聚酶链式反应 温度梯度凝胶电泳(PCR TGGE)和构建16SrRNA基因克隆文库两种方法对比研究了大港油田孔二北断块注水井和采油井的微生物群落结构。16SrDNAV3区PCR扩增产物的TGGE图谱分析表明,这两个油井的微生物群落结构差异很大。注水井样品的TGGE图谱中有6条主要条带,而采油井样品中只有一个条带占绝对优势。同时,建立了两个样品的16SrRNA基因克隆文库,从中分别挑选了10 8和5 0个克隆进行限制性酶切片段长度多样性分析(ARDRA)。注水井样品有33个操作分类单元(OTU) ,其中6个OTU是优势类型;而采油井样品只有8个OTU ,有1个OTU在文库中占绝对优势。克隆文库和TGGE的研究结果一致,均表明注水井样品的微生物多样性比采油井丰富很多。每个OTU的代表克隆序列分析结果表明,注水井样品中的细菌主要属于α、β、γ变形菌纲和放线菌纲,尤其是红细菌亚纲( 4 7% )。采油井样品的细菌主要属于α、β、γ变形菌纲,尤其是假单胞菌属( 6 2 % )。 展开更多
关键词 油藏 16S RRNA基因 TGGE 克隆文库 微生物群落 多样性
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接种微生物菌剂对猪粪堆肥过程中细菌群落多样性的影响 被引量:37
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作者 解开治 徐培智 +5 位作者 张发宝 陈建生 唐拴虎 黄旭 严超 顾文杰 《应用生态学报》 CAS CSCD 北大核心 2009年第8期2012-2018,共7页
利用PCR-DGGE方法研究了接种外源微生物菌剂对鲜猪粪高温好氧堆肥过程中细菌群落多样性的影响.结果表明:接种外源微生物菌剂可以促进堆肥的顺利进行,比不接种处理的高温期提前2d.DGGE图谱分析表明,堆肥中优势细菌群落组成发生了明显的... 利用PCR-DGGE方法研究了接种外源微生物菌剂对鲜猪粪高温好氧堆肥过程中细菌群落多样性的影响.结果表明:接种外源微生物菌剂可以促进堆肥的顺利进行,比不接种处理的高温期提前2d.DGGE图谱分析表明,堆肥中优势细菌群落组成发生了明显的更迭现象,不同堆肥时期细菌群落的Shannon-Wiener指数呈显著差异.目的条带克隆测序结果表明,整个堆肥过程Clostridium stercorariumsub sp.thermolacticum sp.一直是优势菌属,不经培养细菌、Bacillus coagulans sp.、Clostridium thermocellumsp.在接种外源微生物菌剂处理的第10、16天成为优势菌属,不经培养的Firmicutes sp.和不经培养的deltaproteobacterium分别在未接种外源微生物菌剂处理堆肥发酵的第5天和第16天成为优势菌属.非优势菌属Ureibacillus thermosphaericus、不经培养的Silvimonas sp.出现在堆肥腐熟后期,不经培养的土壤细菌主要出现在堆肥初期和高温初期.UPGMC聚类分析表明,接种外源微生物菌剂明显影响了堆肥不同时期的细菌群落结构组成.堆肥化过程中细菌DGGE图谱主成分分析表明,细菌群落变化主要受外源接种微生物菌剂的影响. 展开更多
关键词 微生物菌剂 猪粪 好氧堆肥 细菌 群落多样性 变性梯度凝胶电泳(DGGE)
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堆肥化过程中微生物群落的动态 被引量:35
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作者 王伟东 王小芬 +3 位作者 朴哲 刘长莉 高秀芝 崔宗均 《环境科学》 EI CAS CSCD 北大核心 2007年第11期2591-2597,共7页
通过变性梯度凝胶电泳(DGGE)和平板计数法对堆肥化过程中微生物的区系动态变化进行了研究.结果表明,在堆肥化过程中,微生物数量总的趋势是细菌的数量最多,放线菌次之,真菌的数量最少,中温微生物的数量始终高于高温微生物.当发酵结束后... 通过变性梯度凝胶电泳(DGGE)和平板计数法对堆肥化过程中微生物的区系动态变化进行了研究.结果表明,在堆肥化过程中,微生物数量总的趋势是细菌的数量最多,放线菌次之,真菌的数量最少,中温微生物的数量始终高于高温微生物.当发酵结束后,中温微生物的数量低于发酵初始水平,高温放线菌和高温真菌的数量试验结束后高于初始水平,高温细菌的数量在整个堆肥化过程中变化不大.通过DGGE分析表明,发酵过程中细菌的种类发生了明显的更迭现象.发酵初期Bdellovibrio、Clostridiabacterium、Bacillus、Clostridium等占优势,中期Beta proteobacterium、Petrobacter succinimandens、Nitrospiraebacterium、Clostridium等占优势,后期Clostridium、Beta proteobacterium、Paenibacillus等占优势,而在整个堆肥化过程中Clostridium都是优势种. 展开更多
关键词 堆肥化 微生物区系 变性梯度凝胶电泳
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