Summary of Study ST001805

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001140. The data can be accessed directly via it's Project DOI: 10.21228/M8BQ3C This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

Study ID	ST001805
Study Title	Metabolic responses of two pioneer wood decay fungi to diurnally cycling temperature
Study Summary	1. Decomposition of lignin-rich wood by fungi drives nutrient recycling in woodland ecosystems. Fluctuating abiotic conditions are known to promote the functioning of ecological communities and ecosystems. In the context of wood decay, fluctuating temperature increases decomposition rates. Metabolomics, in tandem with other ‘omics tools, can highlight the metabolic processes affected by experimental treatments, even in the absence of genome sequences and annotations. Globally, natural wood decay communities are dominated by the phylum Basidiomycota. We examined the metabolic responses of Mucidula mucida, a dominant constituent of pioneer communities in beech branches in British woodlands, and Exidia glandulosa, a stress-selected constituent of the same communities, in response to constant and diurnally cycling temperature. 2. We applied untargeted metabolomics and proteomics to beech wood blocks, colonised by M. mucida or E. glandulosa and exposed to either diurnally cycling (mean 15 ± 10°C) or constant (15°C) temperature, in a fully factorial design. 3. Metabolites and proteins linked to lignin breakdown, the citric acid cycle, pentose phosphate pathway, carbohydrate metabolism, fatty acid metabolism and protein biosynthesis and turnover were under-enriched in fluctuating, compared to stable temperatures, in the generalist M. mucida. Conversely E. glandulosa showed little differential response to the experimental treatments. 4. Synthesis. By demonstrating temperature dependant metabolic signatures related to nutrient acquisition in a generalist wood decay fungus, we provide new insights into how abiotic conditions can affect community-mediated decomposition and carbon turnover in forests. We show that mechanisms underpinning important biogeochemical processes can be highlighted using untargeted metabolomics and proteomics in the absence of well-annotated genomes.
Institute	Swansea University
Department	Biosciences
Laboratory	Fungal Molecular Ecology
Last Name	Eastwood
First Name	Daniel
Address	Wallace 102, Biosciences, College of Science, Swansea University, Swansea, SA2 8PP
Email	d.c.eastwood@swansea.ac.uk
Phone	01792513003
Submit Date	2021-05-17
Num Groups	4
Total Subjects	2
Raw Data Available	Yes
Raw Data File Type(s)	d
Analysis Type Detail	GC-MS
Release Date	2021-06-02
Release Version	1

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Sample Preparation:

Sampleprep ID:	SP001888
Sampleprep Summary:	At the end of the experiment, each block was split into three segments from top to bottom using a sterile chisel. The top and middle sections were quenched in liquid nitrogen, lyophilised and stored at -20°C. From the bottom third of each block, four wood chips (~0.5 mm2) were extracted, one from each quadrant, and placed onto 2% (w/v) MEA. Cultures were incubated at 20°C for 7 – 10 days and mycelial morphology confirmed that no woodblocks had become contaminated during the experiment. One third of the frozen woodblock sections from each treatment group were each weighed and manually chipped, added to ≤ 10 ml acetonitrile : methanol : H2O (2:2:1) and shaken at 180 rpm at room temperature for 1 h. Extracts were filtered through glass wool and Whatman filter paper (no. 1) and centrifuged at 3,500 g for 30 min. The supernatant was dried in vacuo (Eppendorf, Hamburg, Germany) overnight. Dried samples were derivitized by addition of 30 μl methoxylamine hydrochloride (15 mg ml-1 in pyridine; Sigma, UK) followed by incubation at 60°C for 90 min. Subsequently 50 μl MSTFA+ 1% (v/v) TMCS (Thermo Scientific) was added to the sample and incubated at 40°C for 60 min. Derivatized samples were transferred to autosampler vials and 10 μl tetracosane (5 mg ml-1 in heptane; Sigma-Aldrich) added as an internal standard.

Sampleprep ID:

SP001888

Sampleprep Summary:

At the end of the experiment, each block was split into three segments from top to bottom using a sterile chisel. The top and middle sections were quenched in liquid nitrogen, lyophilised and stored at -20°C. From the bottom third of each block, four wood chips (~0.5 mm2) were extracted, one from each quadrant, and placed onto 2% (w/v) MEA. Cultures were incubated at 20°C for 7 – 10 days and mycelial morphology confirmed that no woodblocks had become contaminated during the experiment. One third of the frozen woodblock sections from each treatment group were each weighed and manually chipped, added to ≤ 10 ml acetonitrile : methanol : H2O (2:2:1) and shaken at 180 rpm at room temperature for 1 h. Extracts were filtered through glass wool and Whatman filter paper (no. 1) and centrifuged at 3,500 g for 30 min. The supernatant was dried in vacuo (Eppendorf, Hamburg, Germany) overnight. Dried samples were derivitized by addition of 30 μl methoxylamine hydrochloride (15 mg ml-1 in pyridine; Sigma, UK) followed by incubation at 60°C for 90 min. Subsequently 50 μl MSTFA+ 1% (v/v) TMCS (Thermo Scientific) was added to the sample and incubated at 40°C for 60 min. Derivatized samples were transferred to autosampler vials and 10 μl tetracosane (5 mg ml-1 in heptane; Sigma-Aldrich) added as an internal standard.