四川汉源万顺铅锌矿区蜈蚣草共附生细菌群落结构对重金属的响应特征

为了充分挖掘重金属污染地区植物共附生细菌资源,提高重金属污染土壤生态修复的效率,利用变性梯度凝胶电泳(Denaturing Gradient Gel Electrophoresis,DGGE)技术,对四川省汉源县万顺铅锌矿区蜈蚣草根状茎中共附生细菌多样性和群落结构进行了研究,并分析了共附生细菌群落结构与蜈蚣草根状茎中有效重金属间的关系.结果表明,不同采样点间的蜈蚣草共附生细菌多样性存在明显差异,多样性指数和丰富度指数均表现为尾矿矿口弃渣区选矿区农田.基于DGGE回收条带序列的系统发育显示,蜈蚣草共附生细菌包括芽孢杆菌属(Bacillus)、伯克氏菌属(Burkholderia)、根瘤菌属(Rhizobium)、假单胞菌属(Pseudomonas)、肠杆菌属(Enterobacteria)和弯曲杆菌属(Campylobacter)等细菌类群.相关性分析表明,共附生细菌多样性指数和丰富度指数均与有效Cu、Cd、Pb、Zn和As含量呈显著正相关(p0.05).冗余分析(Redundancy Analysis,RDA)显示,有效As是影响蜈蚣草共附生细菌群落结构的关键因子,对砷含量较高的采样点,砷对蜈蚣草共附生细菌群落有促进作用,在砷含量相对较低的采样点,共附生细菌群落与砷呈负相关.研究表明,蜈蚣草存在丰富的共附生细菌种群,这些种群受蜈蚣草中有效重金属含量的影响明显,在修复重金属特别是砷污染土壤方面表现出潜在应用价值.     

尾矿废水对河流沉积物和稻田土壤细菌多样性的影响

采用PCR-DGGE以及分子克隆技术,探讨富含重金属的尾矿废水污染条件下稻田土壤以及河流沉积物中细菌群落结构和多样性变化情况.结果表明,铜尾矿库废水导致了河流沉积物(H1～H6)和稻田土壤(D5)的Fe、As、Cu、Pb和zn污染,其含量远高于未受铜尾矿废水污染的稻田土壤(D1～D4),且As、Fe、Cu的含量显著影响...     

某铁矿尾矿库及周边土壤重金属污染评价

在对某铁矿尾矿库及周边土壤(含水系沉积物、农田表层土壤)重金属含量和理化性质分析测试的基础上,着重分析土壤中重金属元素分布特征,同时运用地累积指数方法对研究区域重金属污染程度进行评价。重金属污染评价结果显示:尾矿库内重金属污染程度排序为CdAsCuZnPbNiCr,元素Cd、As、Cu、Zn均达到严重污染级别;水系沉积物重金属污染程度排序为CdCuAsZnPbNiCr,元素Cd、Cu、As达到严重污染级别;农田表层土壤污染程度CdCuAsZnPbNCr,元素Cd达到严重污染,Cu、As总体虽为中度污染,但个别样点达到重度污染。尾矿库对周边土壤的影响不容忽视,需加强重金属污染的预防和治理。     

铁矿区重金属污染对土壤微生物群落变化的影响

以密云水库上游某铁矿区为研究对象,采用荧光定量PCR研究了矿区内不同采样点土壤中细菌、真菌、放线菌基因数量的变化,并结合土壤的理化性质与重金属污染情况进行了相关分析. 结果表明,研究区微生物的基因数量与该地区的w(AP)(AP为有效磷)均呈显著正相关(P<0.05);细菌、放线菌的基因数量与该地区的内梅罗污染指数呈显著负相关〔二者R分别为-0.756(P<0.01)和-0.614(P<0.05)〕,而真菌的基因数量与内梅罗污染指数无显著相关性. 采用PCR-DGGE研究了细菌、真菌、放线菌的群落结构变化,冗余度分析结果表明,内梅罗污染指数对细菌、放线菌的种群分布影响较大(P<0.05),对真菌的影响则较小. 细菌、放线菌的种群多样性水平随着污染程度升高呈先升后降的趋势. 微生物数量和结构的变化都表明,不同类群的微生物对重金属敏感程度为真菌抗性最强,放线菌和细菌次之. Cd和Cu污染抑制了土壤中微生物的数量和群落多样性,而轻度Cr、Pb和Zn污染则促进了群落数量的增加和群落结构多样性的丰富.      

酸性矿山废水对稻田土壤微生物菌群结构的影响

为探究酸性矿山废水(AMD)对稻田土壤微生物群落结构的影响,通过取自矿区受AMD污染和未受污染的稻田土壤进行微宇宙灌溉模拟实验,研究了AMD污染过程中土壤理化性质和微生物群落的变化,同时建立环境条件变化引起土壤微生物群落结构改变的相关性关系.结果表明,受AMD污染的土壤中SO_4~(2-)、Cd、Zn含量显著上升,土壤酸化且土壤中细菌群落的多样性下降;而恢复清洁水灌溉可提高土壤细菌群落的多样性,有利于修复AMD的污染.采用高通量测序技术分析了不同处理稻田土壤中微生物群落在门和属分类水平上的相对丰度分布变化,冗余分析(RDA)表明,土壤pH和重金属(Pb、Cu)含量是影响稻田土壤微生物群落结构的主要环境因子.研究结果不仅有助于进一步揭示AMD污染、土壤因子与土壤微生物群落的相互关系,同时可为恢复AMD污染农业土壤提供理论依据.     

某铜矿尾矿库对周围环境影响污染评价研究

运用模糊综合型评价法对某铜矿尾矿库区土壤和水体中Cu、Zn、Pb、Cd、Cr 5种重金属含量进行评价以及尾矿库矿渣泥土重金属化学形态分析,结果表明:铜矿尾矿库对周边土壤和水体有不同程度的重金属污染,矿库矿渣泥土为重污染V类,其污染主导因子为Cu;尾矿库周边植被覆盖土为中污染Ⅳ类,其主导污染因子为Cd;矿库渗漏水下游土壤和远离尾矿库区土壤为轻污染Ⅲ类,其主导污染因子为Cd;矿渣泥土重金属各形态以铁锰氧化物结合态、残渣态、可交换态为主,其中Cu的可交换态所占比列极小,为0.24%;尾矿积水、尾矿库渗漏水和尾矿库渗漏水下游溪水均为极严重污染5级,尾矿积水中主导污染因子为Cu、Pb、Cd,尾矿库渗漏水和尾矿库渗漏水下游溪水主导污染因子为Cd。     

钒钛磁铁矿尾矿库复垦土地及周边土壤-玉米重金属迁移富集特征

选取承德中部典型钒钛磁铁矿集中区为研究区,分尾矿库复垦土地、尾矿库周边农用地和对照区农用地系统采集了土壤样品和玉米样品各86件,尾矿砂样品50件,矿石和矿石围岩样品33件.在分析土壤和玉米籽实样品Fe、Cu、Ni、Cd、Cr、Pb、Zn、Hg、Ti、Mn和Mo元素含量和土壤、尾矿砂和矿石围岩重金属赋存形态基础上,采用描述性统计、地累积指数、RDA分析和生物富集系数等方法论述了钒钛磁铁矿尾矿库复垦土地及周边土壤和玉米重金属迁移富集特征.结果表明,尾矿库复垦土地及周边土壤元素地累积程度由强至弱为：P > Cu > Fe₂O₃ > Cr > Ti > V > Ni > Mn > Cd > Zn > Mo > As > Pb > Hg,重金属地累积程度总体属无累积-中度累积水平,土壤Cr和Cu元素超农用地土壤污染筛选值比例为2.32%和1.16%.尾矿库及周边地区玉米籽实Fe、Ti、As、Pb和Mn元素相对较高,对照区玉米籽实Mo、Ni、Cu、Zn、Cd和Cr元素含量相对较高.玉米籽实Pb、As和Ni含量超国家食品限量标准比例分别为18.60%、11.63%和1.16%,均位于尾矿库及其周边区域.尾矿库复垦土地玉米籽实As、Pb、Cd、Cr、Ti和Mn元素生物富集强度相对最高,对照区玉米籽实Cu、Zn和Mo生物富集程度相对较高.尾矿库及周边土壤重金属生物有效态占比Cd元素最高,达50.17%;其次为Ni、Zn和Cu,平均占比为13.61%、13.23%和5.17%.与对照区土壤样品相比,尾矿库复垦土地土壤样品Pb、As和Hg元素表现为总量低,但生物有效性含量和土壤pH高;Cu和Zn总量高但生物有效含量和pH较低特点,使得尾矿库及其周边土壤As和Pb生物富集强度相对增大.在研究矿区尾矿库复垦土地和矿区周边土壤重金属污染生态风险和修复目标值时,宜以重金属生物有效态限值为评价标准.     

中条山十八河铜尾矿库微生物群落组成与环境适应性

为了揭示尾矿区重金属污染与微生物群落组成和多样性的关系,本文选择中条山十八河铜尾矿库的水体、底泥以及矿砂为研究对象,比较分析不同生境中微生物群落组成的差异,及其与环境因子的相关性.结果表明,矿砂中微生物群落的丰富度和多样性最大,而水体中微生物群落的丰富度和多样性最小;在门和目水平上均表现出水体与底泥中微生物群落组成相似,而与矿砂的差异较大;8个优势科中只有鞘脂单胞菌科(Sphingomonadaceae)细菌的相对丰度与重金属(Cd、Cu、Mn、Ni、Pb、Zn)含量存在显著的正相关关系,而其余7个优势科:柄杆菌科(Caulobacteraceae)、甲基杆菌科(Methylobacteriaceae)、类诺卡氏菌科(Nocardioidaceae)、微杆菌科(Microbacteriaceae)、微球菌科(Micrococcaceae)、链球菌科(Streptococcaceae)和类芽孢杆菌科(Paenibacillaceae)细菌的相对丰度与重金属含量存在显著的负相关关系,表明多数细菌都会受到重金属的抑制,而鞘脂单胞菌科细菌对重金属有较高的耐受性,因此具有修复重金属污染的潜力.     

铁矿区内重金属对土壤氨氧化微生物群落组成的影响

以密云水库上游某铁矿区为研究对象,采用荧光定量PCR和变性梯度凝胶电泳(DGGE)分析了矿区内不同采样点的土壤中氨氧化微生物的数量和群落结构的变化,结果表明,土样中氨氧化细菌(AOB)和氨氧化古菌(AOA)的数量变化范围分别为3.01×107~1.08×109copies/g干土和8.65×107~2.69×109copies/g干土.重金属含量与氨氧化微生物数量的相关性分析以及氨氧化微生物群落结构的冗余分析结果表明,该矿区内重金属污染改变了土壤中的氨氧化微生物的数量和结构.Cu污染对AOA的数量起到了显著抑制作用(r= -0.653*, P<0.05),但是对AOB则没有明显作用;Zn污染对尾矿库区域土壤的AOA/AOB比值影响显著(r= -0.606*, P<0.05);Cd污染改变了AOB的种群分布,降低了AOB的多样性水平.土壤中Cr长期干扰并没有改变氨氧化微生物的数量和结构,但是明显得抑制了氨氧化速率,表明重金属污染在一定程度上也影响了土壤生态系统的氮循环.     

三峡澎溪河回水区消落带岸边土壤重金属污染分布特征

在对澎溪河回水区消落带及岸边土样品中重金属含量和样品理化性质测定的基础上,重点分析了该区域内重金属分布特征,并对重金属元素间的相关性展开研究.同时,应用地累积指数对研究区域污染现状进行评价.结果表明,消落带样品中Cu、Cr、Zn、As、Cd、Pb、Hg的平均含量分别为28.17、59.21、108.98、4.77、2.02、28.85、0.52mg·kg-1;岸边土样品中重金属的含量范围分别为22.32、54.90、98.05、7.87、0.77、22.97、0.94mg·kg-1.Cd是三峡库区污染较严重的重金属元素.相关性分析表明：在消落带样品中,Cd与Zn显著相关（p〈0.01）,Pb、Hg和Cu、As都存在显著的正相关关系,说明这4种重金属元素在接受外来污染时可能存在相似性;在岸边土样品中,Cd与Zn、Cr与Cu、As与Hg显著相关（p〈0.01）,Pb与Cu、Cr、Zn、Cd显著正相关,表明这几种重金属可能有着相似的来源.消落带样品重金属污染程度评价结果为：Cd〉Hg〉Zn〉Pb〉Cu〉As〉Cr,岸边土样品重金属污染程度评价结果为：Hg〉Cd〉Zn〉As〉Pb〉Cu〉Cr,Cd和Hg在个别站位达到了严重污染水平.消落带土壤受人为扰动后会成为水体的二次污染源,因此,消落带土壤重金属对水体的潜在影响不容忽视.     

Bacterial diversity in soils around a lead and zinc mine

Five samples of soil collected from a lead and zinc mine were used to assess the effect of combined contamination of heavy metals on soil bacterial communities using a polyphasic approach including characterization of isolates by culture method, community level catabolic profiling in BIOLOG GN microplates, and genetic community fingerprinting by denaturing gradient gel electrophoresis of 16S rDNA fragments amplified by PCR from community DNA （PCR-DGGE）. The structure of the bacterial community was affected to a certain extent by heavy metals. The PCR-DGGE analysis of 16S rRNA genes showed that there were significant differences in the structure of the microbial community among the soil samples, which were related to the contamination levels. The number of bacteria and the number of denaturing gradient gel electrophoresis （DGGE） bands in the soils increased with increasing distance from the lead and zinc mine tailing, whereas the concentration of lead （Pb） and cadmium （Cd） was decreased. Heavily polluted soils could be characterized by a community that differs from those of lightly polluted soils in richness and structure of dominating bacterial populations. The clustering analysis of the DGGE profiles showed that the bacteria in all the five samples of soil belonged to three clusters. The data from the BIOLOG analysis also showed the same result. This study showed that heavy metal contamination decreased both the biomass and diversity of the bacterial community in soil.     

Diversity of free-living nitrogen-fixing microorganisms in wastelands of copper mine tailings during the process of natural ecological restoration

Biological nitrogen fixing is an important source of nitrogen input in the natural ecological restoration of mine wastelands. The diversity of nifH genes in tailings samples under di erent plant communities in Yangshanchong and Tongguanshan wastelands in Tongling, was analyzed using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) approach. The nitrogen-fixing microorganism community in the upper layer of tailings of Tongguanshan wasteland discarded in 1980 showed higher Shannon-Wiener diversity index than that in Yangshanchong wasteland discarded in 1991. The diversity of nifH genes in Yangshanchong wasteland of copper mine tailings did not display a consistent successional tendency with development of plant communities during the process of natural ecological restoration. Phylogenetic analysis of 25 sequences of nifH gene fragments retrieved from the DGGE gels indicated that there were mainly two taxa of free-living nitrogen-fixing microorganisms, Proteobacteria and Cyanobacteria living in the wastelands investigated, most of which were unique and uncultured. Canonical correspondence analysis (CCA) based on the relationship between band patterns of DGGE profile and physico-chemical properties of tailings samples showed that the diversity of nifH genes in di erent tailing samples was mainly a ected by loss of ignition, water content, pH and available Zn contents of wastelands. The dominant plant species and development period of plant communities by ameliorating pH, reducing the toxicity of heavy metals, increasing organic matter and water content a ected the diversity and structure of the free-living nitrogenfixing microorganisms in wastelands of copper mine tailings.     

Detection, isolation, and identification of cadmium-resistant bacteria based on PCR-DGGE

This study focused on the screening of cadmium-resistant bacterial strains from Pb-Zn tailing. We investigated the diversity of microbial community inhabiting Dong-san-cha Pb-Zn tailing in Beijing, China, by polymerase chain reaction-denaturing gradient gel electrophoresis （DGGE） of 16S rRNA gene of bacterial strain, and found two dominant strains in the DGGE profile. Using special culture media, we isolated two strong cadmium-resistant bacterial strains. On the basis of morphological, physiological, and biochemical characteristics, BIOLOG, and 16S rDNA sequencing, the two strains were identified as Bacillus cereus and Enterobacter cloacae. Minimal inhibitory concentrations （MICs） of heavy metals for the bacteria were determined. E. cloacae showed higher MIC values for heavy metals and a larger range of antibiotic resistance than B. cereus.     

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.     

Biolog和PCR-DGGE技术解析椒江口沉积物微生物多样性

采用Biolog和PCR-DGGE技术对椒江口6个站位表层沉积物群落水平的微生物代谢功能和以16S rRNA基因标记的细菌遗传多样性进行分析,并对其代谢功能和群落结构与沉积物污染物等参数进行冗余梯度分析(Redundancy gradient analysis,RDA)和典范对应分析(Canonicalcorrespondence analysis,CCA).Biolog分析结果表明,微生物群落整体代谢活性由高到低的顺序为:潮间带(B1、B2)、入海口处(A3)>入海口内(A1、A2)>入海口外(A4);碳源代谢的Shannon-Wiener多样性指数范围为2.09～3.25,由高到低的顺序为:潮间带(B1、B2)、入海口处(A3)>河道内(A1)>近入海口处(A2)>入海口外(A4);潮间带、入海口处及河道内的微生物对各类碳源的相对利用率较平均,而近入海口处和入海口外的微生物群落对聚合物的相对利用率较高,对氨基酸类和胺类的相对利用率较低.DGGE图谱分析表明,细菌群落结构沿河口盐度梯度存在空间异质性,但两潮间带站位的相似度高(82.27%);遗传基因的Shannon-Wiener多样性指数范围为1.68～2.87,由高到低的顺序为:潮间带>入海口处>入海口外>近入海口处>河道内.群落代谢功能与理化因子的RDA显示,有机质和硝基苯的分布能较好地解释微生物群落代谢功能的变化;群落遗传结构与理化因子的CCA显示,硝基苯和多环芳烃的分布能较好地解释细菌群落遗传结构的变化.综上结果认为,椒江口沉积物的微生物代谢及遗传多样性符合典型的河口特征,但入海口内微生物沉积环境已表现出对某些化工污染物的响应.     

布吉河丰水期总细菌和氨氧化细菌的定性和定量研究

河流中微生物的数量和群落结构能在一定程度上反映水环境状况.氨氧化细菌驱动的硝化作用是氮素转化的主要机制,为了解氮素污染河流中氨氧化细菌的群落组成及数量,采用变性梯度凝胶电泳(denaturing gradient gel electrophoresis,DGGE)和Real-time PCR技术分析了布吉河丰水期不同断面水样中总细菌和氨氧化细菌的群落结构以及数量变化.结果表明,水样中总细菌(16S rRNA)和氨氧化细菌(16S rRNA)数量变化范围分别为4.73×1010～3.90×1011copies.L-1和5.44×106～5.96×108copies.L-1.冗余度分析表明影响微生物数量和群落结构的水环境因子不同:对于总细菌,与其数量显著相关的环境因子是硝氮(P<0.05),与其群落结构显著相关的环境因子是氮素(三氮)和金属(Mn和Zn)(P<0.05);对于氨氧化细菌(ammonia-oxidizing bacteria,AOB),与其数量显著相关的是氨氮和Zn(P<0.05),与其群落组成显著相关的是氨氮、Mn和Zn(P<0.05).测序结果表明在布吉河水样中微生物属于变形菌门(Proteobacterium)的Epsilon-Proteobacteria、Gamma-Proteobacteria、Beta-Proteobacteria和Delta-Proteobacteria这4个纲,氨氧化细菌与Nitrosomonas sp.和Nitrosospira sp.属的细菌相似度较高,且Nitrosospira sp.为优势菌属.由于污染影响,布吉河上游和下游微生物群落结构明显不同.