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首页> 外文期刊>Applied Microbiology >Gene Discovery for Enzymes Involved in Limonene Modification or Utilization by the Mountain Pine Beetle-Associated Pathogen Grosmannia clavigera
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Gene Discovery for Enzymes Involved in Limonene Modification or Utilization by the Mountain Pine Beetle-Associated Pathogen Grosmannia clavigera

机译：山松甲虫相关病原体罗汉果的柠檬烯修饰或利用相关酶的基因发现

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摘要

To successfully colonize and eventually kill pine trees, Grosmannia clavigera (Gs cryptic species), the main fungal pathogen associated with the mountain pine beetle ( Dendroctonus ponderosae ), has developed multiple mechanisms to overcome host tree chemical defenses, of which terpenoids are a major component. In addition to a monoterpene efflux system mediated by a recently discovered ABC transporter, Gs has genes that are highly induced by monoterpenes and that encode enzymes that modify or utilize monoterpenes [especially (+)-limonene]. We showed that pine-inhabiting Ophiostomale fungi are tolerant to monoterpenes, but only a few, including Gs, are known to utilize monoterpenes as a carbon source. Gas chromatography-mass spectrometry (GC-MS) revealed that Gs can modify (+)-limonene through various oxygenation pathways, producing carvone, p -mentha-2,8-dienol, perillyl alcohol, and isopiperitenol. It can also degrade (+)-limonene through the C-1-oxygenated pathway, producing limonene-1,2-diol as the most abundant intermediate. Transcriptome sequencing (RNA-seq) data indicated that Gs may utilize limonene 1,2-diol through beta-oxidation and then valine and tricarboxylic acid (TCA) metabolic pathways. The data also suggested that at least two gene clusters, located in genome contigs 108 and 161, were highly induced by monoterpenes and may be involved in monoterpene degradation processes. Further, gene knockouts indicated that limonene degradation required two distinct Baeyer-Villiger monooxygenases (BVMOs), an epoxide hydrolase and an enoyl coenzyme A (enoyl-CoA) hydratase. Our work provides information on enzyme-mediated limonene utilization or modification and a more comprehensive understanding of the interaction between an economically important fungal pathogen and its host's defense chemicals.
机译：为了成功地定居并最终杀死松树，与山地甲虫（Dendroctonus积木）相关的主要真菌病原体-格罗夫尼亚（Gs隐性物种）已经开发出多种机制来克服寄主树的化学防御，其中萜类是主要成分。除了由最近发现的ABC转运蛋白介导的单萜外排系统外，Gs还具有被单萜高度诱导的基因，其编码修饰或利用单萜的酶[特别是（+）-柠檬烯]。我们表明，居住在松树中的蛇眼雄性真菌对单萜具有耐受性，但是只有少数几个（包括Gs）可以利用单萜作为碳源。气相色谱-质谱（GC-MS）分析表明，Gs可以通过各种氧化途径修饰（+）-柠檬烯，生成香芹酮，对-mentha-2,8-二烯醇，紫苏醇和异胡椒醇。它还可以通过C-1-氧化途径降解（+）-柠檬烯，生成柠檬烯-1,2-二醇作为最丰富的中间体。转录组测序（RNA-seq）数据表明，Gs可能通过β-氧化作用利用柠檬烯1,2-二醇，然后利用缬氨酸和三羧酸（TCA）代谢途径。数据还表明，位于单基因组重叠群108和161中的至少两个基因簇被单萜高度诱导，并可能参与单萜降解过程。此外，基因敲除表明柠檬烯降解需要两种不同的拜尔-维利格单加氧酶（BVMOs），环氧水解酶和烯酰辅酶A（烯酰-CoA）水合酶。我们的工作提供有关酶介导的柠檬烯利用或修饰的信息，以及对经济上重要的真菌病原体与其宿主防御化学物质之间相互作用的更全面的了解。

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