利用T-RFLP解析生物强化去除吡啶过程中微生物种群动态变化

在接种活性污泥处理含吡啶废水的序批式反应器中,引入吡啶降解菌Paracoccus sp.KT-5构成生物强化反应器,研究了对吡啶的生物强化去除特性及效果,利用末端限制性片段长度多态性(T-RFLP)手段解析了微生物种群结构的动态变化.结果表明,投加高效降解菌株KT-5可以加速反应器的启动,但随着反应器的运行,当吡啶初始浓度增加至195.6 mg·L^-1以后,生物强化反应器对吡啶降解的促进作用已不再明显;当吡啶初始浓度在293.4~586.8 mg·L^-1变化时,起初强化反应器对吡啶的去除速率出现了波动,尽管随后逐渐恢复,但仍然没有表现出明显的强化作用.T-RFLP的分析结果表明,当吡啶初始浓度达到978 mg·L^-1以后,生物强化反应器中已检测不到KT-5,表明生物强化作用的消失可能是因为引入的高效降解菌株KT-5的流失造成的.     

Using T-RFLP technology to analyze bacterial diversity in the rhizospheric soils of tea tree at different ages北大核心CSCD

The rhizospheric soils of Tieguanyin at different ages were used as the study materials, and terminal-restriction fragment length polymorphism (T-RFLP) was used to analyze the changes in bacterial diversity. The results showed that the number of T-RFs, Simpson index, and Shannon index decreased significantly with the age of the tea tree. Results of correlation analysis showed that 9 T-RFs from the bacterial community were significantly and positively correlated with the age of the tea tree, and included 34 species of microbes belonging to 10 classes. The 34 microbes were divided into 6 types according to their functional attributes, and included pathogenic bacteria, bacteria that improved soil texture or inhibited pathogenic bacteria, and bacteria associated with the carbon, nitrogen, or sulfur cycles, in which the percentage of pathogenic bacteria was 58.82%. Seventeen T-RFs were significantly and negatively correlated with the age of the tea tree, and included 38 species of bacteria belonging to 12 classes. The 38 bacterial species were divided into 5 types according to their functional attributes, and included pathogenic bacteria, bacteria that improved soil quality or inhibited pathogenic bacteria, and bacteria associated with the carbon cycle or nitrogen cycle, in which all the others, barring the pathogenic bacteria, accounted for 78.95% of the bacterial population. In brief, the diversity and function of bacteria in the rhizospheric soil of tea tree changed significantly with the age of the tea tree, which provides a theoretical basis for studying the interactions of bacterial communities. © 2018 Science Press. All rights reserved.     

Mercury methylation by a microbial community from sediments of the Adour Estuary (Bay of Biscay, France)

In order to study the influence of microorganisms on the mercury biogeochemistry, the metal content and the structure of microbial communities were determined in sediments from stations along the Adour Estuary. The comparison of the bacterial communities and their distribution in function of the environmental parameters by Canonical Correspondence Analysis (CCA) revealed the influence of metals on the bacterial communities structure. Sediments where the bacterial communities are mostly influenced by methylmercury were incubated in slurries with or without mercury, under oxic and anoxic conditions. Methylmercury production was detected in the anoxic biotic slurries with a net methylation yield of 0.3% after 24 h. CCA based on T-RFLP profiles revealed the impact of mercury addition on the bacterial communities structure. In addition, 17 bacterial strains, mainly sulphate-reducing bacteria involved in mercury methylation, were isolated and identified.     

Molecular approaches to microbiological monitoring: fecal source detection

Molecular methods are useful both to monitor natural communities of bacteria, and to track specific bacterial markers in complex environments. Length-heterogeneity polymerase chain reaction (LH-PCR) and terminal restriction fragment length polymorphism (T-RFLP) of 16S rDNAs discriminate among 16S rRNA genes based on length polymorphisms of their PCR products. With these methods, we developed an alternative indicator that distinguishes the source of fecal pollution in water. We amplify 16S rRNA gene fragments from the fecal anaerobic genus Bacteroides with specific primers. Because Bacteroides normally resides in gut habitats, its presence in water indicates fecal pollution. Molecular detection circumvents the complexities of growing anaerobic bacteria. We identified Bacteroides LH-PCR and T-RFLP ribosomal DNA markers unique to either ruminant or human feces. The same unique fecal markers were recovered from polluted natural waters. We cloned and sequenced the unique markers; marker sequences were used to design specific PCR primers that reliably distinguish human from ruminant sources of fecal contamination. Primers for more species are under development. This approach is more sensitive than fecal coliform assays, is comparable in complexity to standard food safety and public health diagnostic tests, and lends itself to automation and high-throughput. Thus molecular genetic markers for fecal anaerobic bacteria hold promise for monitoring bacterial pollution and water quality.     

黄河三角洲湿地土壤微生物群落结构分析

采用末端限制性片段长度多态性分析(T-RFLP)技术和16S rDNA克隆文库的方法,分析了黄河三角洲滨海湿地土壤不同深度细菌和古菌的群落结构.研究表明,随着深度的增加,细菌群落的多样性下降,而古菌群落多样性则有上升的趋势,且土壤的细菌和古菌群落结构都呈现出规律的层状分布.该土壤包括各种硫酸盐还原菌、产甲烷古菌、光合细菌等丰富的细菌和古菌资源.图5参27     

The rhizosphere and PAH amendment mediate impacts on functional and structural bacterial diversity in sandy peat soil

To reveal the degradation capacity of bacteria in PAH polluted soil and rhizosphere we combined bacterial extradiol ring-cleavage dioxygenase and 16S rRNA analysis in Betula pubescens rhizoremediation. Characterisation of the functional bacterial community by RFLP revealed novel environmental dioxygenases, and their putative hosts were studied by 16S rRNA amplification. Plant rhizosphere and PAH amendment effects were detected by the RFLP/T-RFLP analysis. Functional species richness increased in the birch rhizosphere and PAH amendment impacted the compositional diversity of the dioxygenases and the structural 16S rRNA community. A shift from an Acidobacteria and Verrucomicrobia dominated to an Alpha- and Betaproteobacteria dominated community structure was detected in polluted soil. Clone sequence analysis indicated catabolic significance of Burkholderia in PAH polluted soil. These results advance our understanding of rhizoremediation and unveil the extent of uncharacterized functional bacteria to benefit bioremediation by facilitating the development of the molecular tool box to monitor bacterial populations in biodegradation.     

好氧颗粒污泥和活性污泥细菌种群结构对五氯酚污染的响应研究

应用一种新的微生物分子生态学方法--末端限制性片段长度多态性(T-RFLP)法研究了好氧颗粒污泥和活性污泥在毒性化合物五氯酚(PCP)影响下的废水处理性能及其微生物种群结构的响应.结果表明,PCP浓度为30 mg·L-1时,颗粒污泥和活性污泥的COD去除率为38%、77%,与10～20 mg·L-1 PCP相比,分别下降了56%和15%.另外,PCP浓度为20 mg·L-1时,去除率为13%和58%,与10～20 mg·L-1 PCP相比,分别下降了86%和40%,说明PCP对氨氮去除率的影响大于对COD去除率的影响,对颗粒污泥的影响大于对活性污泥的影响.PCP对好氧颗粒污泥和活性污泥的细菌种群结构都产生了明显的影响,而且好氧颗粒污泥的变化程度大于活性污泥;PCP浓度为30 mg·L-1时,好氧颗粒污泥中的细菌种群数量明显下降,TRFs片段数从26下降到14,但活性污泥中的细菌种群数量基本不变,污泥中细菌种群结构的变化趋势与污泥的水处理性能的变化趋势相一致.     

南京8个湖泊超微真核浮游生物遗传多样性的研究

用末端限制性片段长度多态性(terminal restriction fragment length poly morphism,T-RFLP)方法对南京市区8个不同营养水平湖泊的沿岸带和敞水带超微真核浮游生物(0.2～5.0μm)遗传多样性进行了研究,目的是了解不同营养水平湖泊超微真核浮游生物遗传多样性的差异以及影响这种差异的主要因子.T-RFLP结果表明,不同湖泊超微真核浮游生物的T-RFLP指纹图谱存在明显差异,沿岸带和敞水带末端限制性片段T-RFs平均值分别为16.4和15.9,其中营养水平中等的南湖沿岸带T-RFs最多(30个),营养水平较低的百家湖敞水带T-RFs最少(10个),除琵琶湖和莫愁湖外,沿岸带的超微真核浮游生物遗传多样性均比敞水带高.聚类分析表明,除百家湖、前湖、南湖外,其他湖泊沿岸带和敞水带的相似度都比较高.超微真核浮游生物遗传多样性与环境因子的典型对应分析(canonical correspondence analysis,CCA)表明,叶绿素a浓度与超微真核浮游生物群落结构和多样性显著相关(p=0.004).本研究表明,超微真核浮游生物遗传多样性受湖泊营养水平的影响,而且在敞水带和沿岸带存在差异.     

内蒙古高原干涸湖泊反硝化及甲烷氧化细菌的群落分析

以内蒙古高原半干旱草原区干涸湖泊为研究对象,应用末端限制性片段长度多态性(T-RFLP)技术对湿地土壤中的反硝化和甲烷氧化关联菌群的关键功能基因nosZ和pmoA进行了研究,根据末端限制性片段(T-RFs)分析了两种菌群的群落结构及多样性变化,以及环境因素所产生的影响,并探讨了土壤微生物群落对湖泊干涸的响应机制.结果表明:干涸湖泊的湖心、湖底土壤反硝化和甲烷氧化细菌多样性指数较高,群落结构较复杂;而较早干涸的湖坡以及相邻草原对照区,两个菌群多样性指数较低,群落结构相对单一.从湖心到相邻草原,反硝化和甲烷氧化细菌的群落相似度逐渐降低,显示两个菌群处于动态演替中.湖泊干涸过程中,水分含量和有机质含量对反硝化和甲烷氧化细菌的群落结构影响显著;高pH值和高铵态氮分别对反硝化和甲烷氧化细菌有抑制作用.随着湖泊干涸,土壤微生物群落因环境扰动而受到胁迫,这一过程会加剧CO2等温室气体的排放.     

利用T-RFLP技术对温榆河微生物群落结构研究

利用T-RFLP技术(综合运用PCR技术、DNA限制性酶切技术和DNA序列自动分析技术)对温榆河不同尺度、不同时期的微生物群落结构进行了研究.从微生物的种类数、物种丰度、优势菌落、群落结构稳定性等方面特征着手,找出其优势群落;利用两种多样性指数(香侬一威勒多样性指数、辛普森多样性指数)探讨各种微生物群落结构的多样性以及同一群落随时间而发生的变化;比较了微生物指标与水质指标COD之间的变化关系.结果表明,温榆河不同尺度、不同时期的微生物具有明显变化,说明季节性变化对水体中微生物群落产生了明显的影响;T-RF67bp、151bp、440bp、481bp、488bp所代表的微生物群落体现其为优势种群; T-RF为135bp、151bp、440bp的微生物群落对于外界反映最敏感,验证其有可能能作为环境变化的指示性生物;微生物指标与COD呈现明显的正相关.