Summary of Study ST004232

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 PR002670. The data can be accessed directly via it's Project DOI: 10.21228/M8HZ7V 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.

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Study IDST004232
Study TitleConstitutive metabolite profiling of European and Asian Fraxinus with varying susceptibility to ash dieback
Study SummaryHymenoscyphus fraxineus is an invasive pathogen native to East Asia, responsible for the widespread mortality of European ash (Fraxinus excelsior) throughout Europe. Asian ash species, which co-evolved with H. fraxineus, are considered more tolerant than European ash. However, within European ash populations, a small proportion of genotypes show low susceptibility to the pathogen. This study sought to characterize the underlying defence mechanisms to H. fraxineus by performing untargeted constitutive metabolomics profiling of phloem and leaf tissue of susceptible and tolerant European ash and three Asian ash species. Here we report 57 and 36 compounds associated with lower or higher disease susceptibility, from phloem and leaf tissue, respectively. Flavonoids and coumarins were the main classes of detected compounds. In particular, quercitrin and fraxetin exhibited greater variation among the groups. In phloem tissue, quercitrin and fraxetin were more abundant in tolerant than in susceptible European ash and, lowest in Asian ash species. In leaves, however, quercitrin was highest in Asian ash, followed by tolerant and then susceptible European ash. Other flavonoids, coumarins, and iridoid glycosides also showed variation among groups, with stronger differences in phloem than in leaves. Overall, this study advances our understanding of metabolite composition in Fraxinus species with different co-evolutionary histories and susceptibility to H. fraxineus and demonstrates the potential of untargeted metabolomics for investigating defence-related mechanisms in plant-pathogen interactions.
Institute
Swedish University of Agricultural Sciences
DepartmentSouthern Swedish Forest Research Centre
Last NameTolio
First NameBeatrice
AddressSundsvägen 3, 234 56 Alnarp, Sweden
Emailbeatrice.tolio@skogforsk.se; beatrice.tolio@slu.se
Phonena
Submit Date2025-09-09
Raw Data AvailableYes
Raw Data File Type(s)d, mzML
Analysis Type DetailLC-MS
Release Date2025-10-06
Release Version1
Beatrice Tolio Beatrice Tolio
https://dx.doi.org/10.21228/M8HZ7V
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

Select appropriate tab below to view additional metadata details:

Project:

Project ID:PR002670
Project DOI:doi: 10.21228/M8HZ7V
Project Title:Constitutive metabolite profiling of European and Asian Fraxinus with varying susceptibility to ash dieback
Project Summary:Hymenoscyphus fraxineus is an invasive pathogen native to East Asia, responsible for the widespread mortality of European ash (Fraxinus excelsior) throughout Europe. Asian ash species, which co-evolved with H. fraxineus, are considered more tolerant than European ash. However, within European ash populations, a small proportion of genotypes show low susceptibility to the pathogen. This study sought to characterize the underlying defence mechanisms to H. fraxineus by performing untargeted constitutive metabolomics profiling of phloem and leaf tissue of susceptible and tolerant European ash and three Asian ash species. Here we report 57 and 36 compounds associated with lower or higher disease susceptibility, from phloem and leaf tissue, respectively. Flavonoids and coumarins were the main classes of detected compounds. In particular, quercitrin and fraxetin exhibited greater variation among the groups. In phloem tissue, quercitrin and fraxetin were more abundant in tolerant than in susceptible European ash and, lowest in Asian ash species. In leaves, however, quercitrin was highest in Asian ash, followed by tolerant and then susceptible European ash. Other flavonoids, coumarins, and iridoid glycosides also showed variation among groups, with stronger differences in phloem than in leaves. Overall, this study advances our understanding of metabolite composition in Fraxinus species with different co-evolutionary histories and susceptibility to H. fraxineus and demonstrates the potential of untargeted metabolomics for investigating defence-related mechanisms in plant-pathogen interactions.
Institute:University of Copenhagen
Last Name:Crocoll
First Name:Christoph
Address:Thorvaldsensvej 40, Frederiksberg, Hovedstaden, 1871, Denmark
Email:chcr@plen.ku.dk
Phone:+45 35 33 33 69
Funding Source:The study was financially supported by Sveaskog within the framework of the project aimed at ash and elm preservation, Sveriges lantbruksuniversitet Skogsskadecentrum, Kungl. Skogs och Lantbruksakademien (KSLA) grant number CF2022-0009, Partnerskap Alnarp, Stiftelsen för Strategisk Forskning, Stiftelsen fonden för skogsvetenskaplig forskning, Tranemåla stiftelsen, and Extensus stiftelsen.
Publications:Constitutive metabolite profiling of European and Asian Fraxinus with varying susceptibility to ash dieback
Contributors:Tolio, Beatrice; Sherwood, Patrick; Marčiulynienė, Diana; Crocoll, Christoph; Cleary, Michelle; Liziniewicz, Mateusz

Subject:

Subject ID:SU004384
Subject Type:Plant
Subject Species:Fraxinus excelsior, Fraxinus chinensis, Fraxinus mandshurica, Fraxinus platypoda
Taxonomy ID:38873, 56033, 56029, 56030

Factors:

Subject type: Plant; Subject species: Fraxinus excelsior, Fraxinus chinensis, Fraxinus mandshurica, Fraxinus platypoda (Factor headings shown in green)

mb_sample_id local_sample_id Sample source Genotype ID Species Treatment
SA48619328-3001.61LLeaves 3001 Fraxinus excelsior Susceptible European ash
SA48619426-3001.1LLeaves 3001 Fraxinus excelsior Susceptible European ash
SA48619527-3001.3LLeaves 3001 Fraxinus excelsior Susceptible European ash
SA48619629-3001.62LLeaves 3001 Fraxinus excelsior Susceptible European ash
SA48619708-3003.5LLeaves 3003 Fraxinus excelsior Tolerant European ash
SA48619807-3003.2LLeaves 3003 Fraxinus excelsior Tolerant European ash
SA48619906-3003.1LLeaves 3003 Fraxinus excelsior Tolerant European ash
SA48620030-3006.9LLeaves 3006 Fraxinus excelsior Susceptible European ash
SA48620131-3006.1LLeaves 3006 Fraxinus excelsior Susceptible European ash
SA48620232-3006.11LLeaves 3006 Fraxinus excelsior Susceptible European ash
SA48620311-3014.7LLeaves 3014 Fraxinus excelsior Tolerant European ash
SA48620409-3014.1LLeaves 3014 Fraxinus excelsior Tolerant European ash
SA48620510-3014.5LLeaves 3014 Fraxinus excelsior Tolerant European ash
SA48620613-3019.2LLeaves 3019 Fraxinus excelsior Tolerant European ash
SA48620712-3019.1LLeaves 3019 Fraxinus excelsior Tolerant European ash
SA48620816-3024.9LLeaves 3024 Fraxinus excelsior Tolerant European ash
SA48620915-3024.5LLeaves 3024 Fraxinus excelsior Tolerant European ash
SA48621014-3024.4LLeaves 3024 Fraxinus excelsior Tolerant European ash
SA48621134-3025.12LLeaves 3025 Fraxinus excelsior Susceptible European ash
SA48621233-3025.3LLeaves 3025 Fraxinus excelsior Susceptible European ash
SA48621318-3026.12LLeaves 3026 Fraxinus excelsior Tolerant European ash
SA48621417-3026.11LLeaves 3026 Fraxinus excelsior Tolerant European ash
SA48621519-3034.11LLeaves 3034 Fraxinus excelsior Tolerant European ash
SA48621620-3045.6LLeaves 3045 Fraxinus excelsior Tolerant European ash
SA48621736-3049.12LLeaves 3049 Fraxinus excelsior Susceptible European ash
SA48621835-3049.8LLeaves 3049 Fraxinus excelsior Susceptible European ash
SA48621923-3050.9LLeaves 3050 Fraxinus excelsior Tolerant European ash
SA48622022-3050.7LLeaves 3050 Fraxinus excelsior Tolerant European ash
SA48622121-3050.1LLeaves 3050 Fraxinus excelsior Tolerant European ash
SA48622224-3050.1LLeaves 3050 Fraxinus excelsior Tolerant European ash
SA48622325-3050.11LLeaves 3050 Fraxinus excelsior Tolerant European ash
SA48622437-3052.5LLeaves 3052 Fraxinus chinensis Tolerant Asian ash
SA48622538-3052.6LLeaves 3052 Fraxinus chinensis Tolerant Asian ash
SA48622639-3053.2LLeaves 3053 Fraxinus mandshurica Tolerant Asian ash
SA48622740-3053.61LLeaves 3053 Fraxinus mandshurica Tolerant Asian ash
SA48622841-3053.62LLeaves 3053 Fraxinus mandshurica Tolerant Asian ash
SA48622942-3054.7LLeaves 3054 Fraxinus mandshurica Tolerant Asian ash
SA48623043-3055.1LLeaves 3055 Fraxinus platypoda Tolerant Asian ash
SA48623144-3055.3LLeaves 3055 Fraxinus platypoda Tolerant Asian ash
SA48623245-3055.5LLeaves 3055 Fraxinus platypoda Tolerant Asian ash
SA48623346-3056.6LLeaves 3056 Fraxinus platypoda Tolerant Asian ash
SA48623402-57.3aLLeaves 57 Fraxinus excelsior Tolerant European ash
SA48623503-57.3bLLeaves 57 Fraxinus excelsior Tolerant European ash
SA48623604-57.4LLeaves 57 Fraxinus excelsior Tolerant European ash
SA48623705-57.6LLeaves 57 Fraxinus excelsior Tolerant European ash
SA48623801-57.1LLeaves 57 Fraxinus excelsior Tolerant European ash
SA486239QC mix 12Mix of all samples all all -
SA486240QC mix 01Mix of all samples all all -
SA486241QC mix 21Mix of all samples all all -
SA486242QC mix 20Mix of all samples all all -
SA486243QC mix 19Mix of all samples all all -
SA486244QC mix 18Mix of all samples all all -
SA486245QC mix 17Mix of all samples all all -
SA486246QC mix 16Mix of all samples all all -
SA486247QC mix 15Mix of all samples all all -
SA486248QC mix 14Mix of all samples all all -
SA486249QC mix 02Mix of all samples all all -
SA486250QC mix 03Mix of all samples all all -
SA486251QC mix 04Mix of all samples all all -
SA486252QC mix 05Mix of all samples all all -
SA486253QC mix 06Mix of all samples all all -
SA486254QC mix 07Mix of all samples all all -
SA486255QC mix 08Mix of all samples all all -
SA486256QC mix 09Mix of all samples all all -
SA486257QC mix 10Mix of all samples all all -
SA486258QC mix 11Mix of all samples all all -
SA486259QC mix 13Mix of all samples all all -
SA486260QC leaves 02Mix of leaf samples all all -
SA486261QC leaves 01Mix of leaf samples all all -
SA486262QC leaves 04Mix of leaf samples all all -
SA486263QC leaves 05Mix of leaf samples all all -
SA486264QC leaves 03Mix of leaf samples all all -
SA486265QC phloem 01Mix of phloem samples all all -
SA486266QC phloem 05Mix of phloem samples all all -
SA486267QC phloem 04Mix of phloem samples all all -
SA486268QC phloem 03Mix of phloem samples all all -
SA486269QC phloem 02Mix of phloem samples all all -
SA48627029-3001.62PPhloem 3001 Fraxinus excelsior Susceptible European ash
SA48627128-3001.61PPhloem 3001 Fraxinus excelsior Susceptible European ash
SA48627226-3001.1PPhloem 3001 Fraxinus excelsior Susceptible European ash
SA48627327-3001.3PPhloem 3001 Fraxinus excelsior Susceptible European ash
SA48627407-3003.5PPhloem 3003 Fraxinus excelsior Tolerant European ash
SA48627506-3003.2PPhloem 3003 Fraxinus excelsior Tolerant European ash
SA48627608-3014.1PPhloem 3003 Fraxinus excelsior Tolerant European ash
SA48627731-3006.1PPhloem 3006 Fraxinus excelsior Susceptible European ash
SA48627832-3006.11PPhloem 3006 Fraxinus excelsior Susceptible European ash
SA48627930-3006.9PPhloem 3006 Fraxinus excelsior Susceptible European ash
SA48628011-3019.1PPhloem 3014 Fraxinus excelsior Tolerant European ash
SA48628109-3014.5PPhloem 3014 Fraxinus excelsior Tolerant European ash
SA48628210-3014.7PPhloem 3014 Fraxinus excelsior Tolerant European ash
SA48628312-3019.2PPhloem 3019 Fraxinus excelsior Tolerant European ash
SA48628413-3024.3PPhloem 3019 Fraxinus excelsior Tolerant European ash
SA48628514-3024.4PPhloem 3024 Fraxinus excelsior Tolerant European ash
SA48628615-3024.5PPhloem 3024 Fraxinus excelsior Tolerant European ash
SA48628716-3024.9PPhloem 3024 Fraxinus excelsior Tolerant European ash
SA48628833-3025.3PPhloem 3025 Fraxinus excelsior Susceptible European ash
SA48628934-3025.12PPhloem 3025 Fraxinus excelsior Susceptible European ash
SA48629018-3026.12PPhloem 3026 Fraxinus excelsior Tolerant European ash
SA48629117-3026.11PPhloem 3026 Fraxinus excelsior Tolerant European ash
SA48629219-3034.11PPhloem 3034 Fraxinus excelsior Tolerant European ash
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Collection:

Collection ID:CO004377
Collection Summary:The material for this study was selected from a clonal field trial established in 2016 in Snogeholm, Sjöbo, Sweden (55°32'57.4"N 13°42'29.4"E). The trial consisted of 65 replicated ash genotypes: 56 tolerant F. excelsior trees selected from around Sweden, four known susceptible F. excelsior genotypes, and five genotypes representing three Fraxinus species of Asian origin. The Asian ash include: F. mandshurica native to China, Japan, Korea, Eastern Russia; F. platypoda native to China and Japan and, F. chinensis native to northern China, Korea, Japan, south-east Russia (Wallander, 2013). Visually healthy leaf and shoot samples (including bark and phloem) were collected in June 2020 from 13 F. excelsior genotypes varying in disease severity to H. fraxineus (9 tolerant-ET and 4 susceptible-ES) based on periodic assessments, and from all five resistant Asian ash genotypes (AT) (Table S1). Up to five ramets (replicates) per genotype were collected depending on the amount of available tissue. A total of 92 samples were collected in the field, 46 leaves and 46 phloem from the current year shoots. Collected samples were placed in plastic bags and kept on dry ice then stored at -80°C until further processing.
Collection Protocol Filename:Sampling024112.pdf
Sample Type:Plant
Collection Location:55°32'57.4"N 13°42'29.4"E

Treatment:

Treatment ID:TR004393
Treatment Summary:No treatment. This is a genotype study.
Treatment Protocol Filename:Sampling024112.pdf

Sample Preparation:

Sampleprep ID:SP004390
Sampleprep Summary:Samples and tools were kept frozen in liquid nitrogen during all processing steps to minimize degradation or tissue oxidation. Bark including phloem was peeled from the twigs with a sterile scalpel. Bark and leaf tissue were ground separately in liquid nitrogen with porcelain mortars and pestles. For each sample, 100 mg of ground tissue was lyophilized for 24 hours. Metabolites extraction was performed by incubating the tissue with 500 μL of high-performance liquid chromatography grade MeOH containing schaftoside (Phytolab item no. 8332, Vestenbergsgreuth, Germany) as an internal standard at 5 μM. Samples were centrifuged (4000 g, 2 min), and the supernatant was transferred to a new 2 mL microcentrifuge tube and stored at -20°C. The pellet was re-extracted as described above and the new supernatant was combined with previous extraction to get approximately 1000 μL of extract total per sample.

Chromatography:

Chromatography ID:CH005350
Chromatography Summary:Samples were 5-fold diluted with 75% MeOH in water and subjected to analysis by ultra performance liquid chromatography coupled to tandem mass spectrometry (UPLC- MS/MS). UPLC-MS/MS analysis was performed on a Dionex UltiMate 3000 Quaternary Rapid Separation UHPLC+ focused system (Thermo Fisher Scientific, Germering, Germany. many). Separation was achieved on a Kinetex 1.7 μm XB-C18 column (100 × 2.1 mm, 1.7 μm, 100A).
Instrument Name:Thermo Dionex Ultimate 3000 RS
Column Name:Phenomenex Kinetex C18 (100 x 2.1mm,1.7um)
Column Temperature:30 °C
Flow Gradient:0.0−1.0 min 3% B; 1.0−40.0 min 3−30% B; 40.0−50.0 min 30−60% B, 50.0−53.0 min 60−100% B, 53.0−56.0 min 100% B, 56.0−56.5 min 100−3% B, and 56.5−60.0 min 3% B
Flow Rate:300 µl/min
Solvent A:100% water; 0.05 % formic acid
Solvent B:100% Acetonitrile; 0.05 % formic acid
Chromatography Type:Reversed phase

Analysis:

Analysis ID:AN007047
Analysis Type:MS
Operator Name:Christoph Crocoll
Detector Type:ToF
Data Format:.d
Chromatography ID:CH005350
Has Mz:1
Has Rt:1
Rt Units:Minutes
Results File:ST004232_AN007047_Results.txt
Units:Peak area
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