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。

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).