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首页> 外文学位 >A Comprehensive Analysis of the Microbial Diversity Associated with Eunicea Fusca and Related Plexauridae Octocorals, and the Isolation of New Natural Products from Selected, Associated Bacteria.
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A Comprehensive Analysis of the Microbial Diversity Associated with Eunicea Fusca and Related Plexauridae Octocorals, and the Isolation of New Natural Products from Selected, Associated Bacteria.

机译：与Eunicea Fusca和相关的丛生的丛枝菌相关的微生物多样性的综合分析，以及从选定的相关细菌中分离出新的天然产物的方法。

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摘要

Marine invertebrates, including octocorals, are a prolific source of marine natural products (MNPs) and are also known to host diverse microbial communities. It is well-established that marine microbes are producers of bioactive secondary metabolites. Accordingly, it is hypothesized that bioactive metabolites extracted from some marine invertebrates may actually be produced by associated microbes rather than the invertebrate host.;This research aimed to characterize the microbial community of E. fusca and related Plexauridae octocorals and to discover novel, bioactive compounds biosynthesized by associated microorganisms. E. fusca was the main focus of this study because its microbial community has not been thoroughly characterized, and it is the sole source of the potent anti-inflammatory diterpenes fuscol, eunicol, and the fuscosides. Investigations of the unexplored E. fusca microbial community could therefore lead to the discovery of novel MNPs from unexplored microbes, as well as provide insights into the biosynthetic source of these diterpenes.;The microbiome of Floridian and Bahamian E. fusca and related Plexauridae octocorals collected in June 2009 were characterized using culture-independent (454-pyrosequencing, denaturing gradient gel electrophoresis, cloning, and species-specific primers) and -dependent (dilution plating) methods. The culture-independent analysis revealed that E. fusca hosted a geographically-heterogeneous bacterial community, where Endozoicomonas relatives dominated the Florida sites, and Mycoplasma relatives dominated at The Bahamas sites. There were, however, a few stable bacterial associates found in all E. fusca at all sites, including relatives of the Endozoicomonas, Mycoplasma , and Oceanospirillales, suggesting that these bacteria may be critical to holobiont functioning, and differences in environmental conditions at the locations may affect the dominant community member. E. fusca's bacterial community also differed from that of closely-related Plexauridae octocorals and the surrounding seawater. All octocorals contained the same clade of dinoflagellate, Symbiodinium B1/B184. However, the fungal and archaeal communities were not consistent between Plexauridae species or location.;In the culture-dependent study, 137 unique bacteria and 11 unique fungi were cultured from the Plexauridae octocorals; 31 of the isolated bacteria were putative novel species based on 16S rDNA analysis (<97% sequence identity of 16S rDNA). Two of the novel species, Endozoicomonas euniceicola sp. nov. and Endozoicomonas gorgoniicola sp. nov., had nearly identical 16S rDNA sequences to the dominant members in the culture-independent bacterial community, providing the extraordinary opportunity to explore the biology and chemistry of these putative octocoral symbionts. These two bacteria were formally characterized, revealing that they were genotypically, phenotypically, and chemotypically different from the three Endozoicomonas spp. cultures previously isolated from other marine invertebrates.;From the culture-dependent library, selected bacteria were fermented and analyzed for MNP production. Under the evaluated fermentation conditions, two bacteria were found to produce novel MNPs. A Labrenzia sp. produced a family of novel fatty acid derivatives, and a Euzebyella sp. produced a novel 2,5-dialkylresorcinol with good gram-positive bioactivity.;In conclusion, Eunicea fusca hosts a geographically heterogeneous and diverse microbial community that differs from that of other closely-related Plexauridae octocorals and the surrounding environment. This microbial community has a varied metabolic repertoire, which can ultimately lead to the discovery of novel, microbial MNPs.
机译：海洋无脊椎动物（包括八齿动物）是海洋天然产物（MNP）的大量来源，并且还拥有各种微生物群落。众所周知，海洋微生物是生物活性次生代谢物的生产者。因此，假设从某些海洋无脊椎动物中提取的生物活性代谢物实际上可能是由相关的微生物而不是无脊椎动物宿主产生的。这项研究旨在表征大肠埃希菌和相关的丛生八叶虫的微生物群落，并发现新颖的生物活性化合物。由相关微生物生物合成。大肠杆菌（E. fusca）是该研究的主要重点，因为其微生物群落尚未得到充分表征，并且它是强效抗炎性二萜萜烯酚，烯醇和岩藻糖苷的唯一来源。因此，对未开发的真双歧杆菌微生物群落的调查可能会导致从未探索的微生物中发现新的MNP，并为这些二萜的生物合成来源提供见解。使用独立于培养的方法（454焦磷酸测序，变性梯度凝胶电泳，克隆和物种特异性引物）和依赖于培养的方法（稀释平板）对2009年6月的特征进行了表征。不依赖于文化的分析表明，大肠埃希菌（E. fusca）拥有一个地理上异质的细菌群落，其中Endozoicomonas亲戚在佛罗里达州占主导地位，而支原体亲属在巴哈马地区占主导地位。但是，在所有地点的所有大肠埃希菌中都发现了一些稳定的细菌伴生体，包括Endozoicomonas，支原体和Oceospirillales的亲属，这表明这些细菌可能对钩虫的功能以及周围环境条件的差异至关重要。可能会影响主要社区成员。大肠埃希氏菌的细菌群落也与亲缘丛生的丛生八叶虫和周围海水的细菌群落不同。所有八爪鱼都含有相同的壬鞭毛类，Symbiodinium B1 / B184。然而，在丛枝菌属的物种或位置之间，真菌和古细菌群落并不一致。在依赖培养的研究中，从八枝丛菌中培养了137种独特细菌和11种独特真菌。基于16S rDNA分析（16S rDNA的序列同一性<97％），其中31种是推定的新物种。两种新物种，Endozoicomonas euniceicola sp。十一月和Endozoicomonas gorgoniicola sp。 nov。，具有与不依赖培养物的细菌群落中的优势成员几乎相同的16S rDNA序列，为探索这些假定的八叶共生体的生物学和化学性质提供了绝佳的机会。对这两种细菌进行了正式表征，表明它们在基因，表型和化学上均不同于三种内生细菌。以前从其他海洋无脊椎动物中分离得到的细菌。在评估的发酵条件下，发现两种细菌可产生新型MNP。 Labrenzia sp。产生了一个新的脂肪酸衍生物家族和一个Euzebyella sp。产生的新型2,5-二烷基间苯二酚具有良好的革兰氏阳性生物活性。;总的来说，Eusnica fusca拥有一个地理上异质多样的微生物群落，不同于其他紧密相关的丛枝丛八叶虫和周围环境。这个微生物群落具有各种各样的代谢库，最终可以导致发现新的微生物MNP。

著录项

  • 作者

    Pike, Rebecca Emily.;

  • 作者单位

    University of Prince Edward Island (Canada).;

  • 授予单位 University of Prince Edward Island (Canada).;
  • 学科 Biology Microbiology.;Biology Oceanography.
  • 学位 Ph.D.
  • 年度 2014
  • 页码 307 p.
  • 总页数 307
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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