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微宇宙模拟溢油条件下沉积物中蓝细菌多样性
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浙江省自然科学基金项目(No. LR14D030001);国家自然科学基金项目(No. 31370151,31422003)


The impact of oil on the diversity of cyanobacteria in the sediments of a laboratory microcosm
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    摘要:

    【目的】在不同浓度原油胁迫下,研究沉积物中原油降解规律及蓝细菌的响应机制,探讨蓝细菌对原油的耐受程度,并揭示蓝细菌种属结构组成的演替规律,为预测和判断溢油污染是否会造成富营养化等次生环境问题提供依据。【方法】建立微宇宙模拟实验体系,设置未添加原油对照组(CK组)及一系列高浓度原油处理组(25、125和250 g/kg干重,分别为LO组、MO组和HO组),通过气相色谱-氢火焰离子化检测器分析微宇宙培养期间原油降解规律。采用蓝细菌16S rRNA基因分析蓝细菌多样性和丰度变化规律,以及蓝细菌各种属的演替情况。【结果】微宇宙培养第31天时,在最低浓度处理组(LO组)沉积物表层出现片状绿色菌苔,而其他处理组和对照组未出现此现象。蓝细菌16S rRNA基因克隆文库结果表明,在溢油条件下所有处理组的蓝细菌多样性均降低,但有一些种属出现富集,如Oscillatoria和Prochlorococcus等。此外,也监测到大量的不可培养种属,预示着沉积物中可能存在新种属的蓝细菌。【结论】蓝细菌对原油具有较高耐受性,25 g/kg浓度原油会对蓝细菌生长产生刺激作用,且可能进一步引起蓝细菌富集。原油会引起蓝细菌多样性降低,但个别种属会发生富集,说明这些种属对原油具有较高耐受性,甚至发挥重要功能,如Oscillatoria具有固氮功能。本研究在分子水平上研究了蓝细菌对原油的响应,对蓝细菌分子生态研究具有重要意义。

    Abstract:

    [Objective] We studied the oil degradation pattern and the responses of cyanobacteria to crude oil at a series of concentrations, analyzed the resistance of cyanobacteria when treated with oil, explored the succession of cyanobacterial phylotypes under oil stress, and to provide scientific basis to judge if secondary environmental problems such as eutrophication would be caused by oil. [Methods] Microcosm experiments including untreated control (CK) and those treated with crude oil at a series of concentrations (25, 125 and 250 g/kg dry weight (wt), designated as, Light oil (LO), Medium oil (MO) and Heavy oil (HO), respectively) were set up in the laboratory. Gas chromatography-flame ionization detector (GC-FID) was adopted to analyze the degradation of crude oil. Cyanobacterial 16S rRNA genes were used to illustrate the diversity and richness of cyanobacteria and the alteration of genus during incubation period. [Results] A green mat appeared on the surfaces of sediments in the LO group on day 31 and didn’t occur in other groups. Clone libraries of cyanobacterial 16S rRNA genes revealed that the diversity of cyanobacteria reduced in the oiled sediments, while few genus such as Oscillatoria and Prochlorococcus increased at later stage. In addition, large proportion of sequences affiliated to uncultured genus might imply novel species. [Conclusion] This study indicated that cyanobacteria exhibited high resistance to crude oil. Oil could stimulate the cyanobacteria growth and might lead to cyanobacteria bloom in some situations. This study circumvents traditional cumbersome technologies, such as isolation and culture-based methods for cyanobacteria investigation, which is significant to molecular ecology analysis on cyanobacteria.

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解翠晓,张凯,孙永革,吕镇梅. 微宇宙模拟溢油条件下沉积物中蓝细菌多样性[J]. 微生物学通报, 2017, 44(7): 1579-1588

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  • 在线发布日期: 2017-06-30
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