玉米秸秆厌氧降解复合菌系的微生物群落结构

以不同来源具有木质纤维素降解能力的环境样品为接种物,以玉米秸秆为唯一碳源,进行了秸秆厌氧降解微生物的驯化培养,构建了8组复合菌系.复合菌系的产气周期为30～50 d,玉米秸秆的总固体(TS)去除率在30%～40%之间.其中6组的甲烷产量(以TS计)为62.1～118.4 mL.g-1,发酵液中挥发性有机酸主要为乙酸、丙酸及丁酸(100～500 mg.L-1),发酵终产物pH为6.5～6.7;而另外2组的甲烷产量较低,在8.5～9.7 mL.g-1之间,但乙酸浓度较高(1 200 mg.L-1左右),发酵终产物pH为5.6～5.9.通过16S rRNA基因克隆文库方法对复合菌系中细菌及古菌的群落结构进行了多样性解析,结果表明细菌主要分为8个不同的类群,其中厚壁菌门(Firmicutes)、拟杆菌门(Bacteroidetes)、互养菌门(Synergistetes)及热袍菌门(Thermotogae)为优势菌群,分别占细菌克隆总数的37.8%、34.3%、11.6%和6.4%;古菌属于甲烷微菌纲(Methanomicrobia)和甲烷杆菌纲(Methanobacteria),分别占古菌克隆总数的61.1%和38.9%.对主要功能菌群在秸秆厌氧降解产甲烷过程中发挥的作用进行了探讨.     

高效氯氰菊酯降解菌群的结构分析及其降解特性研究

以连作10年以上的棉田土壤为材料,以高效氯氰菊酯为唯一碳源,驯化获得能稳定传代并持续降解高效氯氰菊酯的LZ1菌群,对菌群中可培养的细菌进行分离鉴定,最后对其降解高效氯氰菊酯的特性进行分析。结果表明,LZ1菌群中可分离、纯化优势菌株12株,经16SrRNA序列分析,其中10株与铜绿假单胞菌(Pseudomonas aeruginosa)相似性达99%,1株与无色杆菌(Achromobacter mucicolens)相似性达99%。高效氯氰菊酯最佳反应条件为高效氯氰菊酯初始质量浓度250mg/L、温度27℃、pH7.0、装样量200mL。在最佳反应条件下培养的LZ1菌群24h时对高效氯氰菊酯的降解率可达68.81%,132h时的降解率可达92.39%,且LZ1菌群对高效氯氰菊酯的4种异构体没有明显的降解特异性。     

基于16S rRNA基因测序法分析北京霾污染过程中PM2.5和PM10细菌群落特征

2013年1月8~14日,北京出现了严重的霾污染.霾污染时高浓度的大气颗粒物增加了暴露人群的健康风险,而大气中的微生物也可能带来一些风险,但目前对霾污染时大气中微生物组成了解较少.本研究选取了2013年1月8~14日北京7d的PM2.5和PM10采样样本,通过对细菌16S rRNA基因V3区扩增和Mi Seq测序,得到PM2.5和PM10中的细菌群落结构特征,并将结果与相同采样样本的宏基因组测序结果及三项国外基于16S rRNA基因测序方法的大气中细菌研究结果进行了比较.研究发现7 d连续采样条件下,PM2.5中细菌群落结构在门和属的水平上均差别不大.在属级别上,节杆菌属(Arthrobacter)和弗兰克氏菌属(Frankia)是北京冬季大气中细菌群落的主要类群.16S rRNA基因测序与宏基因组测序结果对比分析发现,在属级别上,两种分析方法中有39个相同的属类群(两种分析方法丰度前50的细菌类群合并所得),弗兰克氏菌属(Frankia)和副球菌属(Paracoccus)在16S rRNA基因测序分析结果中相对含量较多,而考克氏菌属(Kocuria)和地嗜皮菌属(Geodermatophilus)在宏基因组测序结果中相对含量较高.在门和属的水平上,PM2.5和PM10中细菌群落结构特征呈现出相似的规律.在门水平上,放线菌门(Actinobacteria)在PM2.5中的相对百分比较大,而厚壁菌门(Firmicutes)在PM10中的相对百分比较大.在属水平上,梭菌属(Clostridium)在PM10中的相对百分比较大.与三项国外基于16S rRNA基因测序研究结果对比发现,尽管在采样地点和采样时间上有较大差异,大气中普遍存在一些相同的细菌类群,且近地面大气细菌群落结构特征相似度较高,区别于高空对流层中细菌群落结构特征.     

九龙江流域潜在病原菌污染分析

近年来,由于流域经济快速发展和城镇化进程加快等原因,九龙江水污染问题日趋严重.为全面认识九龙江流域病原菌的分布状况,应用16S rRNA基因-454焦磷酸测序技术测定九龙江支流西溪、北溪水体和沉积物中细菌16S rRNA基因V1~V3高变区,共获得204 216条高质量序列.通过与病原菌参考数据库对比分析发现,九龙江分布有68个潜在病原菌属,占序列总量的6.1%.其中梭菌属(Clostridium)、分支杆菌属(Mycobacterium)和鞘氨醇单胞菌属(Sphingomonas)在所有样品中都有检出,且丰度最高,分别占病原菌属序列总量的54.5%、5.9%和5.6%.在种水平上,九龙江分布有48种病原菌,占序列总量的0.76%.其中Afipia felis、Mycobacterium asiaticum、Clostridium baratii、Brucella melitensis和Delftia tsuruhatensis是丰度最高的5种病原菌,分别占病原菌种序列总量的48.9%、20.3%、8%、2.7%和1.7%.统计分析表明,九龙江水体中分布的病原菌种类数(属或种)显著高于沉积物中,且西溪水体检出的病原菌种类(属或种)和丰度最高,说明九龙江水体,尤其是西溪水体受病原菌污染的风险较高.此外,相关分析表明,九龙江水体中病原菌的种类数(属或种)和丰度与营养盐(氮、磷)有着显著的正相关关系,说明九龙江水体中分布的病原菌与沿岸的人类活动,如养殖业、污水排放等密切相关.因此,为保证流域的公共卫生安全,需进一步加强污染源清理整顿工作,开展水环境病原菌的实时监测.     

常温低基质下碱度和溶解氧对厌氧氨氧化的影响

通过改变进水碱度、光照条件和溶解氧,对以陶粒为填料的常温低基质上向流厌氧氨氧化生物滤柱的脱氮性能进行了研究.借助克隆测序技术对滤柱内的微生物菌群进行了分析.结果表明,水温为20℃±2℃,水力停留时间为3 h条件下,当进水碱度在44~350 mg·L-1时,随着碱度的降低,氨氮去除率从97.2%降至75.6%,总氮去除率从89.7%降至75.1%,而亚硝氮去除率稳定在99.7%,进水碱度为0 mg·L-1时,出水亚硝氮浓度突跃至4.9 mg·L-1.当进水碱度恢复至350 mg·L-1,存在自然光照情况下,氨氮去除率较没有光照时的氨氮去除率降低12.3%.水力停留时间为1.5 h条件下,当进水DO小于3mg·L-1时,平均氨氮去除率和亚硝氮去除率分别为99.7%和100%,平均总氮去除负荷为1.0 kg·(m3·d)-1.16S rRNA克隆测序鉴定结果表明,滤柱下部的厌氧氨氧化菌种类为Candidatus Jettenia asiatica和Candidatus Brocadia sp.,这两种菌对20℃±2℃水温有很好的适应性.     

Vertical diversity of sediment bacterial communities in two different trophic states of the eutrophic Lake Taihu, China

Vertical diversity of sediment bacterial communities in 2 different trophic states (macrophyte-dominated and algae-dominated) of the large shallow eutrophic Lake Taihu, China, were investigated using denaturing gradient gel electrophoresis (DGGE) and 16S rRNA sequence analysis. Clustering analysis of DGGE profiles showed that different clusters were recognized in different depths of sediment cores in the 2 lake trophic states. Analyses of the bacterial diversity, as estimated by the Shannon index (H'), showed that different sediment layers of the macrophyte-dominated state had higher diversity than the algae-dominated state. In addition, bacterial diversity of the sediment in the macrophyte-dominated state changed abruptly throughout the layers, but bacterial diversity of the algae-dominated state decreased gradually with sediment depth. Phylogenetic analysis showed that Proteobacteria was the most abundant phylum in the middle sediment of the 2 lake trophic states. In the macrophyte-dominated state, clone sequences related to Betaproteobacteria (50.0%) were the most abundant, followed by Epsilonproteobacteria (21.1%), Acidobacteria (7.9%), Deltaproteobacteria (7.9%), Chloroflexi (7.9%), and Bacteroidetes (5.3%); whereas in the algae-dominated state, sequences affiliated with Betaproteobacteria (84.4%) were predominant, followed by Deltaproteobacteria (12.5%) and Acidobacteria (3.1%). Canonical correspondence analysis showed that organic matter and pH play key roles in driving the vertical changes of bacterial community composition.     

Denaturing gradient gel electrophoresis fingerprinting of soil bacteria in the vicinity of the Chinese Great Wall Station, King George Island, Antarctica

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.     

高通量测序技术研究辽河真核浮游藻类的群落结构特征

真核浮游藻类在河流生态系统中发挥着重要生态功能,本研究对辽河水环境样本的18S rRNA基因的V4可变区进行454焦磷酸测序,对获得的OTU代表性序列进行物种注释.比较注释后的生物种类信息与光学显微镜观察结果的异同,并将OTUs的代表性序列与NCBI中真核浮游藻类的18S rRNA基因序列进行系统发育分析,此外研究了真核浮游藻类群落结构特征及其环境影响指标.研究共获得约167 901条18S rRNA基因V4可变区的高质量序列,注释为424个OTUs,涵盖了134种真核浮游藻类OTUs.与光学显微镜鉴别结果类似,高通量测序结果以硅藻门和绿藻门为优势类群,此外检测到轮藻门、隐藻门、定鞭藻门和金藻纲等显微镜观察结果中未有的类群.系统发育分析可在门这一分类阶元对隐藻门和甲藻门聚类,并将隐藻门在属水平进行聚类和区分,甲藻门可以在部分科或属水平聚类和区分.RDA结果显示,氨氮、活性磷和硝态氮是影响真核浮游藻类群落结构特征最重要的环境因子.本研究为认识辽河水环境中真核浮游藻类的多样性、群落结构及其环境影响因子提供了新的视角,同时也预示着高通量测序技术在辽河流域浮游植物监测和水环境质量指示中的应用潜力.     

Determination of growth kinetics of microorganisms linked with 1,4-dioxane degradation in a consortium based on two improved methods

● Evaluated three methods for determining the consortia’s growth kinetics. ● Conventional method is flawed since it relies on the total biomass concentration. ● Considering only selected bacterial taxa improved the accuracy. ● Considering oligotrophs and copiotrophs further improved the accuracy. The conventional method for determining growth kinetics of microbial consortia relies on the total biomass concentration. This may be inaccurate for substrates that are uncommon in nature and can only be degraded by a small portion of the microbial community. 1,4-dioxane, an emerging contaminant, is an example of such substrates. In this work, we evaluated an improved method for determining the growth kinetics of a 1,4-dioxane-degrading microbial consortium. In the improved method, we considered only bacterial taxa whose concentration increase correlated to 1,4-dioxane concentration decrease in duplicate microcosm tests. Using PEST (Parameter Estimation), a model-independent parameter estimator, the kinetic constants were estimated by fitting the Monod kinetics-based simulation results to the experimental data that consisted of the concentrations of 1,4-dioxane and the considered bacterial taxa. The estimated kinetic constants were evaluated by comparing the simulation results with experimental results from another set of microcosm tests. The evaluation was quantified by the sum of squared relative residual, which was four orders of magnitude lower for the improved method than the conventional method. By further dividing the considered bacterial taxa into oligotrophs and copiotrophs, the sum of squared relative residual further decreased.     

不同种植方式对亚热带红壤微生物多样性的影响

土壤微生物在推动土壤碳循环过程方面发挥着重要作用,然而种植方式的改变对土壤微生物多样性的影响机制还不十分清楚.本研究采集湖南省盘塘县长期定位试验站红壤稻田(PR)、旱地(UC)及水旱轮作(PR)这3种不同种植方式的土壤样品,采用末端限制性酶切片段长度多态性分析(T-RFLP)技术和实时荧光定量(RT-PCR)技术分析了土壤细菌16S rRNA基因的多样性和丰度,研究种植方式改变对土壤微生物数量、群落结构及其多样性的影响.结果表明,3种种植方式的土壤细菌16S rRNA基因数量(以干土计)为2.5×109~1.5×1010拷贝·g-1,与PR相比,UP和UC处理16S rRNA基因丰度显著下降(P0.05).同时,3种种植方式下土壤细菌的优势类群为变形菌(76、90和327 bp;相对丰度47%~53%)和绿弯菌(65 bp;相对丰度10%~12%).冗余分析表明种植方式改变了土壤理化性质,导致土壤细菌群落结构特征的显著变化,而土壤理化性质中有机碳和全氮含量是影响土壤细菌群落结构的主要因子.多样性指数分析(香农指数和均匀度指数)显示种植水稻的土壤细菌多样性最高,显著高于水旱轮作和旱地土壤.可见,种植方式的改变对土壤群落组成和数量造成了深刻的影响,而水稻种植是亚热带红壤可持续利用的一种有效方式,其更有利于土壤有机质的累积,土壤肥力及微生物多样性均较高.