厦门近海海水中多环芳烃降解菌的原位富集与降解菌多样性  

In situ enrichment and diversity analysis of polycyclic aromatic carbon degrading bacteria in the coastal seawater of Xiamen Island

作　　者：陈亮[1,2] 董纯明[2] 何进[1] 邵宗泽[2] 

机构地区：[1]华中农业大学生命科学技术学院,农业微生物学国家重点实验室,武汉430070 [2]国家海洋局第三海洋研究所,海洋生物遗传资源重点实验室,厦门361005

出　　处：《微生物学报》2010年第10期1392-1398,共7页Acta Microbiologica Sinica

基　　金：福建省自然科学基金资助项目(2010J01206);福建省科技项目(2009H0029);国家自然科技资源平台项目(2005DKA21209)~~

摘　　要：【目的】为了分析厦门近海原位海水中多环芳烃降解菌的多样性。【方法】将涂有菲的聚氯乙烯(PVC)板悬挂在厦门国际邮轮码头的海水中,进行菲降解菌的原位富集。利用变性梯度凝胶电泳(Denaturinggradient gel electrophoresis,DGGE)和16S rRNA基因文库两种方法分析了在PVC板表面富集微生物的菌群结构。之后,在实验室模拟原位条件下,对PVC板表面富集的菲降解菌群进行进一步富集、分离和初步鉴定。【结果】PVC板在海水中浸没6 d后,16S rRNA基因文库分析表明,在涂菲的PVC板表面富集的菌群中解环菌属(Cycloclasticus)对应的克隆子占文库总克隆子的50%;在未涂菲的PVC板表面吸附的菌群中红杆菌科(Rhodobacteraceae)为优势菌,其对应的克隆子占文库总克隆子的47%;而解环菌属的克隆子只占文库总克隆子的2%。DGGE的分析结果也证明解环菌是菲原位富集降解菌群中的优势菌。实验室进一步富集后,从该菌群中分离鉴定出14株细菌,其中一株新鞘氨醇杆菌B14(Novosphingobium sp.B14)具有菲降解能力。但是,解环菌未能获得纯培养。【结论】菲原位富集发现,厦门近海水体中解环菌是多环芳烃的主要降解菌。[ Objective] The aim of this study is to assess the diversity of polycyclic aromatic carbon （PAH）-degrading bacteria in the coastal seawater of Xiamen Island. [ Methods ] The phenanthrene-degradlng bacteria were enriched by suspending phenanthrene-eoated Polyvinylchloride （PVC） plates in the seawater close to Xiamen International Cruise Dock. PCR-DGGE and 16S rRNA gene clone library were used to analyze the bacteria colonizing on the PVC plates. Further, PAH-degrading bacteria were re-enriched in lab after the in situ enrichment and subjected to diversity analysis and bacterial isolation. [ Results] After 6 days incubation, the genus Cycloclasticus was shown to be the dominant bacterium on the phenanthrene （Phe）-coated plates, which accounted for 50% of the total clones in 16S rRNA gene clone library. However, on the control plates without Phe-coating, bacteria of Rhodobacteraceae were the dominant member, which accounted for 47% of the total clones. PCR-DGGE results reconfirmed the genus Cycloclasticus as the dominant member on the Phe-coated plates. After re-enrichment with Phe in laboratory, 14 strains were isolated from the consortium, which contained a potential novel species of genus Novosphingobium, named strain B-14 and identified as a Phe degrader. However, the most predominant member Cycloclasticus can not be cultivated into pure culture. [ Conclusion] Genus Cycloclasticus is the most important PAH-degrading bacterium in the coast sea water of Xiamen Island. This is the first evidence to our knowledge about the in situ enrichment PAH-degrading bacterium in seawater.

关 键 词：厦门近海 多环芳烃 解环菌 生物降解 原位富集 

分 类 号：X172[环境科学与工程—环境科学]