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

摘要:

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HinfⅠ两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

Abstract:

Abatract [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.