溴代阻燃剂污染底泥与未污染底泥中细菌群落结构之比较分析  

Comparison of bacterial diversity of polluted and unpolluted sediment by Brominated Flame Retardant

作　　者：赵恒芝[1,2] 戴世鲲[1] 孙伟[1,2] 王广华[1] 郭雅涛[1,2] 郭俊[3] 许枚英[3] 吴后波[1] 李翔[1] 

机构地区：[1]中国科学院南海海洋研究所,中国科学院海洋生物可持续利用重点实验室,广州510301 [2]中国科学院研究生院,北京100049 [3]广东省微生物研究所,广州510070

出　　处：《微生物学报》2011年第3期377-385,共9页Acta Microbiologica Sinica

基　　金：广东省自然科学基金项目(9351007002000001);广东省科技计划项目(2008A030203004)~~

摘　　要：【目的】通过免培养的分子生物学方法,比较受溴代阻燃剂污染严重的河流底泥和与未污染水库底泥中细菌多样性差异,解析二者间细菌群落结构,为污染河流的治理与生物修复提供相关的理论依据。【方法】从中国贵屿溴代阻燃剂污染区练江底泥样品和未污染水库底泥样品中分别提取微生物总DNA,用细菌通用引物27F和1500R扩增16S rRNA基因,构建16S rRNA基因文库。用HhaⅠ和HinfⅠ2种限制性内切酶对克隆子进行扩增产物rDNA的限制性酶切分析(ARDRA),挑取不同的操作分类单元OUT中的克隆进行测序并构建系统发育树,比较代表克隆子的基本分类类群和生物多样性构成。【结果】未污染水库底泥中细菌群落组成主要包括变形细菌门(Proteobacteria)的α、β、γ和δ4个亚门、浮霉菌门(Planctomycetes)、酸杆菌门(Acidobacteria)、放线菌门(Actinobacteria)、绿弯菌门(Chloroflexi)、疣微菌门(Verrucomicrobia)和厚壁菌门(Firmicutes)。优势菌是酸杆菌,占文库克隆的30.2%。污染河流底泥中细菌群落组成包括变形细菌门(Proteobacteria)的α、β、δ和ε4个亚门、浮霉菌门(Planctomycetes)、绿菌门或拟杆菌门(Chlorobi orBacteroidetes)、酸杆菌门(Acidobacteria)、放线菌门(Actinobacteria)、绿弯菌门(Chloroflexi)、待定菌群candidatedivision OP01、candidate division OP08和candidate division WS3的相似菌。优势菌是ε-变形细菌和绿弯菌,占文库克隆的44.9%。【结论】受溴代阻燃剂污染河流底泥中的细菌群落具有较高水平的多样性,与未污染底泥有显著区别,主要体现在ε-变形细菌和绿弯菌在细菌群落中具有优势地位。这种优势种群的改变可能与污染物的过度富集具有一定的相关性,对于我们进一步探索和了解溴代阻燃剂的微生物修复具有一定的指导意义。[Objective]By using brominated flame retardant we compared the bacterial diversity of highly polluted river sediment with that of nearby unpolluted lake.[Method]Total DNA was extracted from unpolluted and highly polluted sediment sample by brominated flame retardant in Guiyu of China.The 16S rRNA gene was amplified by PCR using bacterial primer 27F and 1500R.The plasmid libraries of the amplicons were constructed.The positive clones with insert were screened on plates with IPTG/X-gal /Ap.Amplified ribosmal DNA restriction analysis（ARDRA） was carried out with restriction enzymes HhaⅠand HinfⅠ.Representative clones of each operational taxonomic unit based on the ARDRA patterns were selected to be sequenced.After proof reading and careful comparison to remove the chimeric sequences,the partial sequence of 16S rRNA gene were used for construction of the phylogenetic tree.[Result]The result of blast searching showed that clones from unpolluted sediment sample belonged to α-Proteobacteria,β-Proteobacteria,γ-Proteobacteria,δ-Proteobacteria,Planctomycetes,Acidobacteria,Actinobacteria,Chloroflexi,Verrucomicrobia,Firmicutes,the predominant bacteria（30.2% of total clones） is Acidobacteria;most clones from polluted sediment belonged to α-Proteobacteria,β-Proteobacteria,ε-Proteobacteria,δ-Proteobacteria,Planctomycetes,Acidobacteria,Actinobacteria,Chloroflexi,Bacteroidetes,Firmicutes,candidate division OP01,candidate division OP08,the predominant bacteria（44.9% of total clones） are ε-Proteobacteria and Chloroflexi.[Conclusion]Bacterial community structure of polluted sediment has distinguished feature and obviously different from the unpolluted sediment sample,which is mainly reflected in the dominant position of ε-Proteobacteria and Chloroflexi in the bacterial flora.

关 键 词：溴代阻燃剂 细菌群落结构 16S RRNA基因 ARDRA 

分 类 号：X172[环境科学与工程—环境科学]