潮间带沉积物厌氧烃降解细菌的多样性及<i>Desulfovibrio subterraneus</i> ND17的分离鉴定

张琪; 邓晔; 邵宗泽; 王万鹏

doi:10.12284/hyxb2022154

潮间带沉积物厌氧烃降解细菌的多样性及Desulfovibrio subterraneus ND17的分离鉴定

doi: 10.12284/hyxb2022154

张琪^{1, 2,},
邓晔^{1, 3},
邵宗泽²,
王万鹏^2, ,

1.
山东大学海洋研究院，山东青岛 266237
2.
自然资源部第三海洋研究所海洋生物遗传资源重点实验室，福建厦门 361005
3.
中国科学院生态环境研究中心，北京 100085

基金项目: 水圈微生物计划（91851203）；国家自然科学基金（41876143, 41922041）；福建省自然科学基金重点项目（2021J02015）。

详细信息

作者简介:
张琪（1995-），女，山东省德州市人，研究方向为厌氧烃降解微生物。E-mail：1278271178@qq.com

通讯作者:
王万鹏，研究员，研究方向为微生物的石油烃降解机理。E-mail：wangwanpeng@tio.org.cn

中图分类号: Q938.8；Q939.1
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出版历程
- 收稿日期: 2022-02-26
- 修回日期: 2022-04-13
- 网络出版日期: 2022-04-25
- 刊出日期: 2022-08-15

Diversity of anaerobic hydrocarbon-degrading bacteria in intertidal sediments and isolation and identification of Desulfovibrio subterraneus ND17

Zhang Qi^{1, 2
,},
Deng Ye^{1, 3},
Shao Zongze²,
Wang Wanpeng^{2
, ,}

1.
Institute of Marine Science and Technology, Shandong University, Qingdao 266237, China
2.
Key Laboratory of Marine Biogenetic Resources, Third Institute of Oceanography, Ministry of Natural Resources, Xiamen 361005, China
3.
Research Center for Eco-Environmental Sciences, Chinese Academy of Science, Beijing 100085, China

摘要

摘要: 潮间带作为海陆交界处，易受到来自海洋的石油污染，且各类石油烃进入沉积物后的降解过程尚不清楚。前人在各类生境中对好氧微生物烃降解方面已有较多研究，但对近海潮间带环境中的厌氧烃降解鲜有报道。本研究对青岛女岛湾潮间带沉积物深层样品以混合烃（中长链烷烃、多环芳烃）为碳源，硫酸盐作为电子受体进行厌氧富集培养。富集菌群的细菌多样性表明在混合烃作为碳源的作用下，优势菌群转变为脱硫叠球菌科（Desulfosarcinaceae）、脱硫杆菌科（Desulfobacteraceae）等具有石油烃降解潜力的硫酸盐还原菌。经分离纯化得到一株厌氧烃降解菌ND17，与地下脱硫弧菌属模式种Desulfovibrio subterraneus HN2^T 16S rRNA基因序列的相似度为99.93%。进一步实验表明，菌株ND17在厌氧条件下对二十四烷和菲的降解率可分别达到53.9%和35.7%。这也是首次对脱硫弧菌属单菌在厌氧条件下进行石油烃降解的研究。脱硫弧菌作为一种广泛分布在厌氧环境的细菌，本研究为进一步认识其在海洋石油污染环境中的修复潜力提供了支撑。
- 潮间带 /
- 厌氧烃降解 /
- 硫酸盐还原菌 /
- 脱硫弧菌属
Abstract: As a junction between land and sea, the intertidal zone is vulnerable to oil pollution from the ocean, and the degradation process of various types of petroleum hydrocarbons entering the sediment is still unclear. There have been many studies on aerobic microbial hydrocarbon degradation, but little has been reported on anaerobic hydrocarbon degradation in the offshore intertidal environment. In this study, anaerobic enrichment culture was conducted on deep intertidal sediment samples from Qingdao Nüdao Bay with mixed hydrocarbons (medium and long chain alkanes and polycyclic aromatic hydrocarbons) as carbon source and sulfate as electron acceptor. The bacterial diversity of the enriched flora indicated that the dominant flora changed to Desulfosarcinaceae, Desulfobacteraceae and other sulfate-reducing bacteria with petroleum hydrocarbon degradation potential under the effect of mixed hydrocarbons as carbon source. An anaerobic hydrocarbon-degrading strain ND17 was obtained by isolation and purification, with 99.93% similarity to the 16S rRNA gene sequence of Desulfovibrio subterraneus HN2^T, the model species of Desulfovibrio subterraneus genus. Further experiments showed that strain ND17 could degrade eicosanoids and phenanthrene up to 53.9% and 35.7%, respectively, under anaerobic conditions. This is the first study on the degradation of petroleum hydrocarbons under anaerobic conditions by a single bacterium of the genus Desulfovibrio. As a widely distributed bacterium in anaerobic environments, this study supports further understanding of its remediation potential in marine petroleum-contaminated environments.
- intertidal zone /
- anaerobic hydrocarbon degradation /
- sulfate-reducing bacteria /
- Desulfovibrio

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图 1 原位和富集样本的细菌序列稀释曲线

Fig. 1 Rarefaction curves of bacterial sequences for in situ and enriched samples

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图 2 原位和富集样本的细菌Alpha多样性

Fig. 2 Bacterial Alpha diversity for in situ and enriched samples

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图 3 原位和富集样本中不同分类水平的细菌群落组成柱形图

Fig. 3 Barplots of bacterial community composition at different taxonomic levels in in situ and enriched samples

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图 4 基于Weighted UniFrac的PCoA结果

Fig. 4 Results of PCoA based on Weighted UniFrac

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图 5 原位和富集样本组间群落丰度显著性差异检验

Fig. 5 Test for significant differences in community abundance between in situ and enriched sample groups

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图 6 菌株ND17基于16S rRNA基因序列的系统发育分析

Fig. 6 Phylogenetic analysis of strain ND17 based on 16S rRNA gene sequences

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图 7 菌株ND17对菲和二十四烷的降解速率的测定

Fig. 7 Determination of degradation rates of phenanthrene and eicosanoids by strain ND17

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