Microbial lipid cycling in the oceans: an integrated 'omics' approach

海洋中的微生物脂质循环：综合“组学”方法

• 批准号: 2881140
• 金额: --
• 依托单位: University of Warwick
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2881140
• 关键词: Microbial; lipid; cycling; oceans; integrated

Lipids, containing carbon, nitrogen, phosphorus and sulfur, are a major component of all living cells and are constantly released into the oceans due to programmed cell death, viral infections and zooplankton grazing. Current estimates suggest that lipids can account for ~10% of total dissolved organic matter (DOC) in the oceans. However, our knowledge on microbial lipid transformation in the oceans is very limited. Neither the identities of the microbes involved, nor the mechanisms by which lipids are degraded, have been well understood. A major hurdle in understanding the microbial lipid cycle lies in the difficulty in characterizing and quantifying numerous lipid classes in marine samples. Commonly observed lipids in marine ecosystems include phospholipids, glycolipids and amino acid-containing lipids. Recent advances in technologies such as liquid chromatography electrospray ionization mass spectrometry (LC-ESI-MS) has enabled high throughput identification and quantification of thousands of cellular lipid molecular species (Shevchenko, 2010), which now enables lipid characterization both qualitatively and quantitatively. Indeed, lipidomics is starting to complement the rapid progress made in genomics, metagenomics, transcriptomics and proteomics. Here in this project, we will combine laboratory and filed based methodologies to fully uncover the lipid cycle in marine waters. The aim of this project is thus twofold, Firstly, we aim to establish a high throughput workflow for simultaneous determination of common lipids species in marine surface waters using our in house HPLC MS platform (see above figure 1). This will involve examining samples from the Western English Channel, collected via the Marine Biological Association research vessel, Sepia. Secondly, we aim to determine the identity and activity of key microbial populations in the recycling of representative lipids in marine surface waters and coastal marine sediment using integrated omics approach, e.g. high throughput sequencing of 16S/18S rRNA, metagenomics, metagenomics and lipidomics. Microbial lipids identification will be carried out by liquid chromatography-mass spectrometry (LC-ESI-MS). Nutrients and key intermediates in lipid degradation pathways will be analysed by established chromatography techniques, such as ion-exchange chromatography and gas chromatography, and analysis guided by our previous extensive characterisation of laboratory strains. Microbial community analyses will be carried out using amplicon sequencing of 16S rRNA genes, metatranscriptomics and metagenomics.

含有碳、氮、磷和硫的脂质是所有活细胞的主要组成部分，由于细胞程序性死亡、病毒感染和浮游动物摄食，脂质不断释放到海洋中。目前的估计表明，脂质可以占海洋中总溶解有机物（DOC）的约10%。然而，我们对海洋中微生物脂质转化的了解非常有限。无论是所涉及的微生物的身份，还是脂质降解的机制，都还没有得到很好的理解。了解微生物脂质循环的一个主要障碍在于难以表征和定量海洋样品中的众多脂质类。海洋生态系统中常见的脂质包括磷脂、糖脂和含氨基酸的脂质。液相色谱电喷雾电离质谱（LC-ESI-MS）等技术的最新进展已实现了对数千种细胞脂质分子种类的高通量鉴定和定量（Shevchenko，2010），现在可以定性和定量地进行脂质表征。事实上，脂质组学正开始补充基因组学、宏基因组学、转录组学和蛋白质组学的快速进展。在这个项目中，我们将结合联合收割机实验室和现场为基础的方法，以充分揭示海洋沃茨中的脂质循环。因此，该项目的目的是双重的，首先，我们的目标是建立一个高通量的工作流程，使用我们的内部HPLC MS平台同时测定海洋表面沃茨中的常见脂质种类（见上图1）。这将涉及检查通过海洋生物协会研究船Sepia从西英吉利海峡收集的样本。其次，我们的目标是确定在海洋表面沃茨和沿海海洋沉积物中的代表性脂质的回收中的关键微生物种群的身份和活性，使用集成的组学方法，例如16 S/18 S rRNA的高通量测序，宏基因组学，宏基因组学和脂质组学。将通过液相色谱-质谱法（LC-ESI-MS）进行微生物脂质鉴别。脂质降解途径中的营养物质和关键中间体将通过已建立的色谱技术（如离子交换色谱法和气相色谱法）进行分析，并通过我们先前对实验室菌株的广泛表征进行分析。将使用16 S rRNA基因的扩增子测序、元转录组学和元基因组学进行微生物群落分析。