Biodiversity and Hydrocarbon Degradation Capacity of Marine Fungi

我对海洋真菌的生物多样性及其降解碳氢化合物的能力感兴趣。我更感兴趣的是在芬迪湾及其边缘盐沼（加拿大东部）自然生长的真菌种类，以及它们对海洋石油泄漏的自然恢复力。我的实验室正在解决这个问题，首先使用形态学和分子方法（DNA条形码）表征水柱和沿海沉积物中海洋真菌的多样性。然后，我们将使用以船用燃料油为唯一碳源的选择性培养基来分离潜在的烃降解真菌。这些实验将使我们能够评估原油的质量损失，并提供碳氢化合物代谢的证据（C14萘降解为CO2）。它们还将有助于确定哪些海洋真菌拥有细胞色素P450单加氧酶（P450），真菌使用这些酶来降解碳氢化合物。从新型真菌分离株和环境样品中鉴定和定量P450的方法将包括通过液相色谱-轨道阱串联质谱（LC-MS/MS）进行蛋白质鉴定和通过液相色谱选择反应监测质谱（LC-SRM）进行定量。这项工作将与Erin Bertrand博士（达尔豪西第二层CRC海洋微生物蛋白质组学）合作完成。 我实验室的初步工作已经从新斯科舍省的海水中鉴定出了候选菌株，这些菌株使我们能够调查先前暴露于石油是否会增加海洋真菌对石油的降解率。我们还打算研究候选真菌菌株单独和混合种财团，以研究营养补充和温度对石油降解率的影响。我们将使用我们的蛋白质组学方法来探索天然石油降解真菌和现有的芬迪湾微生物群落（抑制或促进降解酶的生产）在不同的温度和营养制度之间的相互作用。 我们正在描述这一鲜为人知的沿海系统组成部分，以探索真菌在芬迪湾生态系统海洋石油泄漏恢复能力中的作用。我的实验室是第一个采用分子生态学方法来了解海洋真菌的生物多样性和石油降解能力的实验室。在这个过程中，我们也将开始揭示海洋真菌的进化关系，改善我们的参考数据库，以DNA为基础的真菌鉴定，并培训学生在尖端的分子生物学和蛋白质组学方法。我们期望描述许多新的真菌物种，为我们阐明海洋真菌的生物多样性和生态功能在北方沿海生态系统。我的工作将有广泛的影响，在冷水海洋生态系统的石油泄漏的补救，并了解真菌细胞色素P450的演变。

I am interested in the biodiversity of marine fungi and in their ability to degrade hydrocarbons. I am more specifically interested in the species of fungi that grow naturally in the Bay of Fundy and its fringing saltmarshes (eastern Canada) and the extent to which they have natural resilience to marine oil spills. My lab is approaching this problem by first characterizing the diversity of marine fungi from the water column and coastal sediments using morphological and molecular methods (DNA barcoding). We will then use selective media with marine fuel oil as sole carbon source to isolate potential hydrocarbon-degrading fungi. These experiments will allow us to assess the mass loss of crude oil, and provide proof of metabolism of hydrocarbons (C14 naphthalene degradation to CO2). They will also help determine which marine fungi possess cytochrome P450 monooxygenases (P450s), which fungi use to degrade hydrocarbons. Approaches for identifying and quantifying P450s from novel fungal isolates and environmental samples will include protein identification via liquid chromatography-orbitrap tandem mass spectrometry (LC-MS/MS) and quantification via liquid chromatography-selected reaction monitoring mass spectrometry (LC-SRM). This work will be done in collaboration with Dr. Erin Bertrand (Dalhousie Tier II CRC in Marine Microbial Proteomics). Preliminary work in my lab has identified candidate strains from Nova Scotia seawater that allow us to investigate if prior exposure to oil increases oil degradation rates by marine fungi. We also intend to study candidate fungal strains singly and in mixed-species consortia to examine the effects of nutrient supplementation and temperature on oil degradation rates. We will use our proteomics approach to explore interactions between native oil degrading fungi and the existing Bay of Fundy microbial community (inhibition or promotion of degradative enzyme production) under different temperature and nutrient regimes. We are characterizing this poorly known component of coastal systems to explore the role that fungi play in the resilience to marine oil spills of the Bay of Fundy ecosystem. My lab is the first to take a molecular ecology approach to understanding the biodiversity and oil degrading capacity of marine fungi. In the process, we will also begin to uncover the evolutionary relationships of marine fungi, improving our reference databases for DNA-based fungal identifications, and training students in cutting edge molecular phylogenetics and proteomics approaches. We expect to describe many new species of fungi as we shed light on marine fungal biodiversity and ecological function in northern coastal ecosystems. My work will have broad implications for remediation of oil spills in coldwater marine ecosystems, and for understanding the evolution of fungal cytochrome P450s.