#METABOLOMICS WORKBENCH ccrocoll_20250909_024334 DATATRACK_ID:6404 STUDY_ID:ST004232 ANALYSIS_ID:AN007047 PROJECT_ID:PR002670
VERSION             	1
CREATED_ON             	September 29, 2025, 7:52 pm
#PROJECT
PR:PROJECT_TITLE                 	Constitutive metabolite profiling of European and Asian Fraxinus with varying
PR:PROJECT_TITLE                 	susceptibility to ash dieback
PR:PROJECT_SUMMARY               	Hymenoscyphus fraxineus is an invasive pathogen native to East Asia, responsible
PR:PROJECT_SUMMARY               	for the widespread mortality of European ash (Fraxinus excelsior) throughout
PR:PROJECT_SUMMARY               	Europe. Asian ash species, which co-evolved with H. fraxineus, are considered
PR:PROJECT_SUMMARY               	more tolerant than European ash. However, within European ash populations, a
PR:PROJECT_SUMMARY               	small proportion of genotypes show low susceptibility to the pathogen. This
PR:PROJECT_SUMMARY               	study sought to characterize the underlying defence mechanisms to H. fraxineus
PR:PROJECT_SUMMARY               	by performing untargeted constitutive metabolomics profiling of phloem and leaf
PR:PROJECT_SUMMARY               	tissue of susceptible and tolerant European ash and three Asian ash species.
PR:PROJECT_SUMMARY               	Here we report 57 and 36 compounds associated with lower or higher disease
PR:PROJECT_SUMMARY               	susceptibility, from phloem and leaf tissue, respectively. Flavonoids and
PR:PROJECT_SUMMARY               	coumarins were the main classes of detected compounds. In particular, quercitrin
PR:PROJECT_SUMMARY               	and fraxetin exhibited greater variation among the groups. In phloem tissue,
PR:PROJECT_SUMMARY               	quercitrin and fraxetin were more abundant in tolerant than in susceptible
PR:PROJECT_SUMMARY               	European ash and, lowest in Asian ash species. In leaves, however, quercitrin
PR:PROJECT_SUMMARY               	was highest in Asian ash, followed by tolerant and then susceptible European
PR:PROJECT_SUMMARY               	ash. Other flavonoids, coumarins, and iridoid glycosides also showed variation
PR:PROJECT_SUMMARY               	among groups, with stronger differences in phloem than in leaves. Overall, this
PR:PROJECT_SUMMARY               	study advances our understanding of metabolite composition in Fraxinus species
PR:PROJECT_SUMMARY               	with different co-evolutionary histories and susceptibility to H. fraxineus and
PR:PROJECT_SUMMARY               	demonstrates the potential of untargeted metabolomics for investigating
PR:PROJECT_SUMMARY               	defence-related mechanisms in plant-pathogen interactions.
PR:INSTITUTE                     	University of Copenhagen
PR:LAST_NAME                     	Crocoll
PR:FIRST_NAME                    	Christoph
PR:ADDRESS                       	Thorvaldsensvej 40, Frederiksberg, Hovedstaden, 1871, Denmark
PR:EMAIL                         	chcr@plen.ku.dk
PR:PHONE                         	+45 35 33 33 69
PR:FUNDING_SOURCE                	The study was financially supported by Sveaskog within the framework of the
PR:FUNDING_SOURCE                	project aimed at ash and elm preservation, Sveriges lantbruksuniversitet
PR:FUNDING_SOURCE                	Skogsskadecentrum, Kungl. Skogs och Lantbruksakademien (KSLA) grant number
PR:FUNDING_SOURCE                	CF2022-0009, Partnerskap Alnarp, Stiftelsen för Strategisk Forskning,
PR:FUNDING_SOURCE                	Stiftelsen fonden för skogsvetenskaplig forskning, Tranemåla stiftelsen, and
PR:FUNDING_SOURCE                	Extensus stiftelsen.
PR:PUBLICATIONS                  	Constitutive metabolite profiling of European and Asian Fraxinus with varying
PR:PUBLICATIONS                  	susceptibility to ash dieback
PR:CONTRIBUTORS                  	Tolio, Beatrice; Sherwood, Patrick; Marčiulynienė, Diana; Crocoll, Christoph;
PR:CONTRIBUTORS                  	Cleary, Michelle; Liziniewicz, Mateusz
#STUDY
ST:STUDY_TITLE                   	Constitutive metabolite profiling of European and Asian Fraxinus with varying
ST:STUDY_TITLE                   	susceptibility to ash dieback
ST:STUDY_SUMMARY                 	Hymenoscyphus fraxineus is an invasive pathogen native to East Asia, responsible
ST:STUDY_SUMMARY                 	for the widespread mortality of European ash (Fraxinus excelsior) throughout
ST:STUDY_SUMMARY                 	Europe. Asian ash species, which co-evolved with H. fraxineus, are considered
ST:STUDY_SUMMARY                 	more tolerant than European ash. However, within European ash populations, a
ST:STUDY_SUMMARY                 	small proportion of genotypes show low susceptibility to the pathogen. This
ST:STUDY_SUMMARY                 	study sought to characterize the underlying defence mechanisms to H. fraxineus
ST:STUDY_SUMMARY                 	by performing untargeted constitutive metabolomics profiling of phloem and leaf
ST:STUDY_SUMMARY                 	tissue of susceptible and tolerant European ash and three Asian ash species.
ST:STUDY_SUMMARY                 	Here we report 57 and 36 compounds associated with lower or higher disease
ST:STUDY_SUMMARY                 	susceptibility, from phloem and leaf tissue, respectively. Flavonoids and
ST:STUDY_SUMMARY                 	coumarins were the main classes of detected compounds. In particular, quercitrin
ST:STUDY_SUMMARY                 	and fraxetin exhibited greater variation among the groups. In phloem tissue,
ST:STUDY_SUMMARY                 	quercitrin and fraxetin were more abundant in tolerant than in susceptible
ST:STUDY_SUMMARY                 	European ash and, lowest in Asian ash species. In leaves, however, quercitrin
ST:STUDY_SUMMARY                 	was highest in Asian ash, followed by tolerant and then susceptible European
ST:STUDY_SUMMARY                 	ash. Other flavonoids, coumarins, and iridoid glycosides also showed variation
ST:STUDY_SUMMARY                 	among groups, with stronger differences in phloem than in leaves. Overall, this
ST:STUDY_SUMMARY                 	study advances our understanding of metabolite composition in Fraxinus species
ST:STUDY_SUMMARY                 	with different co-evolutionary histories and susceptibility to H. fraxineus and
ST:STUDY_SUMMARY                 	demonstrates the potential of untargeted metabolomics for investigating
ST:STUDY_SUMMARY                 	defence-related mechanisms in plant-pathogen interactions.
ST:INSTITUTE                     	Swedish University of Agricultural Sciences
ST:DEPARTMENT                    	Southern Swedish Forest Research Centre
ST:LAST_NAME                     	Tolio
ST:FIRST_NAME                    	Beatrice
ST:ADDRESS                       	Sundsvägen 3, 234 56 Alnarp, Sweden
ST:EMAIL                         	beatrice.tolio@skogforsk.se; beatrice.tolio@slu.se
ST:PHONE                         	na
#SUBJECT
SU:SUBJECT_TYPE                  	Plant
SU:SUBJECT_SPECIES               	Fraxinus excelsior, Fraxinus chinensis, Fraxinus mandshurica, Fraxinus platypoda
SU:TAXONOMY_ID                   	38873, 56033, 56029, 56030
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data
SUBJECT_SAMPLE_FACTORS           	57.1	01-57.1L	Sample source:Leaves | Genotype ID:57 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=01-BT220720-L-01 57.1 5x neg_GA1_01_33955_modified.mzML
SUBJECT_SAMPLE_FACTORS           	57.1	01-57.1P	Sample source:Phloem | Genotype ID:57 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=01-BT220720-P-01 57.1 5x neg_BA1_01_34020_modified.mzML
SUBJECT_SAMPLE_FACTORS           	57.3a	02-57.3aL	Sample source:Leaves | Genotype ID:57 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=02-BT220720-L-02 57.3a 5x neg_GB1_01_33970_modified.mzML
SUBJECT_SAMPLE_FACTORS           	57.3a	02-57.3P	Sample source:Phloem | Genotype ID:57 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=02-BT220720-P-02 57.3 5x neg_BB1_01_34027_modified.mzML
SUBJECT_SAMPLE_FACTORS           	57.3b	03-57.3bL	Sample source:Leaves | Genotype ID:57 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=03-BT220720-L-03 57.3b 5x neg_GC1_01_33961_modified.mzML
SUBJECT_SAMPLE_FACTORS           	57.3b	03-57.4P	Sample source:Phloem | Genotype ID:57 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=03-BT220720-P-03 57.4 5x neg_BC1_01_34005_modified.mzML
SUBJECT_SAMPLE_FACTORS           	57.4	04-57.4L	Sample source:Leaves | Genotype ID:57 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=04-BT220720-L-04 57.4 5x neg_GD1_01_33935_modified.mzML
SUBJECT_SAMPLE_FACTORS           	57.4	04-57.6P	Sample source:Phloem | Genotype ID:57 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=04-BT220720-P-04 57.6 5x neg_BD1_01_34003_modified.mzML
SUBJECT_SAMPLE_FACTORS           	57.6	05-57.6P	Sample source:Phloem | Genotype ID:57 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=05-BT220720-L-05 57.6 5x neg_GE1_01_33938_modified.mzML
SUBJECT_SAMPLE_FACTORS           	57.6	05-57.6L	Sample source:Leaves | Genotype ID:57 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=05-BT220720-P-05 3003.1 5x neg_BE1_01_34041_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3003.1	06-3003.1L	Sample source:Leaves | Genotype ID:3003 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=06-BT220720-L-06 3003.1r5x neg_GF1_01_33986_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3003.1	06-3003.2P	Sample source:Phloem | Genotype ID:3003 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=06-BT220720-P-06 3003.2 5x neg_BF1_01_33999_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3003.2	07-3003.2L	Sample source:Leaves | Genotype ID:3003 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=07-BT220720-L-07 3003.2 5x neg_GG1_01_33972_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3003.2	07-3003.5P	Sample source:Phloem | Genotype ID:3003 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=07-BT220720-P-07 3003.5 5x neg_BG1_01_34022_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3003.5	08-3003.5L	Sample source:Leaves | Genotype ID:3003 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=08-BT220720-L-08 3003.5 5x neg_GH1_01_33963_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3003.5	08-3014.1P	Sample source:Phloem | Genotype ID:3003 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=08-BT220720-P-08 3014.1 5x neg_BH1_01_33993_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3014.1	09-3014.1L	Sample source:Leaves | Genotype ID:3014 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=09-BT220720-L-09 3014.1 5x neg_GA2_01_33979_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3014.1	09-3014.5P	Sample source:Phloem | Genotype ID:3014 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=09-BT220720-P-09 3014.5 5x neg_BA2_01_34015_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3014.5	10-3014.5L	Sample source:Leaves | Genotype ID:3014 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=10-BT220720-L-10 3014.5 5x neg_GB2_01_33957_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3014.5	10-3014.7P	Sample source:Phloem | Genotype ID:3014 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=10-BT220720-P-10 3014.7 5x neg_BB2_01_34031_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3014.7	11-3014.7L	Sample source:Leaves | Genotype ID:3014 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=11-BT220720-L-11 3014.7 5x neg_GC2_01_33944_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3014.7	11-3019.1P	Sample source:Phloem | Genotype ID:3014 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=11-BT220720-P-11 3019.1 5x neg_BC2_01_33995_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3019.1	12-3019.1L	Sample source:Leaves | Genotype ID:3019 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=12-BT220720-L-12 3019.1 5x neg_GD2_01_33975_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3019.1	12-3019.2P	Sample source:Phloem | Genotype ID:3019 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=12-BT220720-P-12 3019.2 5x neg_BD2_01_34037_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3019.2	13-3019.2L	Sample source:Leaves | Genotype ID:3019 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=13-BT220720-L-13 3019.2 5x neg_GE2_01_33964_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3019.2	13-3024.3P	Sample source:Phloem | Genotype ID:3019 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=13-BT220720-P-13 3024.3 5x neg_BE2_01_34034_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3024.4	14-3024.4L	Sample source:Leaves | Genotype ID:3024 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=14-BT220720-L-14 3024.4 5x neg_GF2_01_33934_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3024.4	14-3024.4P	Sample source:Phloem | Genotype ID:3024 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=14-BT220720-P-14 3024.4 5x neg_BF2_01_34038_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3024.5	15-3024.5L	Sample source:Leaves | Genotype ID:3024 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=15-BT220720-L-15 3024.5 5x neg_GG2_01_33933_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3024.5	15-3024.5P	Sample source:Phloem | Genotype ID:3024 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=15-BT220720-P-15 3024.5 5x neg_BG2_01_34036_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3024.9	16-3024.9L	Sample source:Leaves | Genotype ID:3024 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=16-BT220720-L-16 3024r9 5x neg_GH2_01_33987_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3024.9	16-3024.9P	Sample source:Phloem | Genotype ID:3024 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=16-BT220720-P-16 3024.9 5x neg_BH2_01_34013_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3026.11	17-3026.11L	Sample source:Leaves | Genotype ID:3026 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=17-BT220720-L-17 3026.11 5xneg_GA3_01_33950_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3026.11	17-3026.11P	Sample source:Phloem | Genotype ID:3026 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=17-BT220720-P-17 3026.11 5xneg_BA3_01_33996_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3026.12	18-3026.12L	Sample source:Leaves | Genotype ID:3026 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=18-BT220720-L-18 3026.12 5xneg_GB3_01_33966_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3026.12	18-3026.12P	Sample source:Phloem | Genotype ID:3026 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=18-BT220720-P-18 3026.12 5xneg_BB3_01_34009_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3034.11	19-3034.11L	Sample source:Leaves | Genotype ID:3034 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=19-BT220720-L-19 3034.11 5xneg_GC3_01_33977_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3034.11	19-3034.11P	Sample source:Phloem | Genotype ID:3034 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=19-BT220720-P-19 3034.11 5xneg_BC3_01_34016_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3045.6	20-3045.6L	Sample source:Leaves | Genotype ID:3045 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=20-BT220720-L-20 3045.6 5x neg_GD3_01_33965_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3045.6	20-3045.6P	Sample source:Phloem | Genotype ID:3045 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=20-BT220720-P-20 3045.6 5x neg_BD3_01_34014_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3050.1	21-3050.1L	Sample source:Leaves | Genotype ID:3050 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=21-BT220720-L-21 3050.1 5x neg_GE3_01_33985_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3050.1	21-3050.1P	Sample source:Phloem | Genotype ID:3050 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=21-BT220720-P-21 3050.1 5x neg_BE3_01_34040_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3050.7	22-3050.7L	Sample source:Leaves | Genotype ID:3050 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=22-BT220720-L-22 3050.7 5x neg_GF3_01_33948_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3050.7	22-3050.7P	Sample source:Phloem | Genotype ID:3050 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=22-BT220720-P-22 3050.7 5x neg_BF3_01_34000_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3050.9	23-3050.9L	Sample source:Leaves | Genotype ID:3050 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=23-BT220720-L-23 3050.9 5x neg_GG3_01_33945_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3050.9	23-3050.9P	Sample source:Phloem | Genotype ID:3050 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=23-BT220720-P-23 3050.9 5x neg_BG3_01_33998_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3050.1	24-3050.1L	Sample source:Leaves | Genotype ID:3050 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=24-BT220720-L-24 3050.1 5x neg_GH3_01_33984_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3050.1	24-3050.1P	Sample source:Phloem | Genotype ID:3050 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=24-BT220720-P-24 3050.1 5x neg_BH3_01_34023_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3050.11	25-3050.11L	Sample source:Leaves | Genotype ID:3050 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=25-BT220720-L-25 3050.11 5xneg_GA4_01_33954_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3050.11	25-3050.11P	Sample source:Phloem | Genotype ID:3050 | Species:Fraxinus excelsior | Treatment:Tolerant European ash	RAW_FILE_NAME(RAW-FILE_NAME)=25-BT220720-P-25 3050.11 5xneg_BA4_01_34030_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3001.1	26-3001.1L	Sample source:Leaves | Genotype ID:3001 | Species:Fraxinus excelsior | Treatment:Susceptible European ash	RAW_FILE_NAME(RAW-FILE_NAME)=26-BT220720-L-26 3001.1 5x neg_GB4_01_33980_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3001.1	26-3001.1P	Sample source:Phloem | Genotype ID:3001 | Species:Fraxinus excelsior | Treatment:Susceptible European ash	RAW_FILE_NAME(RAW-FILE_NAME)=26-BT220720-P-26 3001.1 5x neg_BB4_01_34033_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3001.3	27-3001.3L	Sample source:Leaves | Genotype ID:3001 | Species:Fraxinus excelsior | Treatment:Susceptible European ash	RAW_FILE_NAME(RAW-FILE_NAME)=27-BT220720-L-27 3001.3 5x neg_GC4_01_33973_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3001.3	27-3001.3P	Sample source:Phloem | Genotype ID:3001 | Species:Fraxinus excelsior | Treatment:Susceptible European ash	RAW_FILE_NAME(RAW-FILE_NAME)=27-BT220720-P-27 3001.3 5x neg_BC4_01_34026_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3001.61	28-3001.61L	Sample source:Leaves | Genotype ID:3001 | Species:Fraxinus excelsior | Treatment:Susceptible European ash	RAW_FILE_NAME(RAW-FILE_NAME)=28-BT220720-L-28 3001.61 5xneg_GD4_01_33983_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3001.61	28-3001.61P	Sample source:Phloem | Genotype ID:3001 | Species:Fraxinus excelsior | Treatment:Susceptible European ash	RAW_FILE_NAME(RAW-FILE_NAME)=28-BT220720-P-28 3001.61 5xneg_BD4_01_33992_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3001.62	29-3001.62L	Sample source:Leaves | Genotype ID:3001 | Species:Fraxinus excelsior | Treatment:Susceptible European ash	RAW_FILE_NAME(RAW-FILE_NAME)=29-BT220720-L-29 3001.62 5xneg_GE4_01_33969_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3001.62	29-3001.62P	Sample source:Phloem | Genotype ID:3001 | Species:Fraxinus excelsior | Treatment:Susceptible European ash	RAW_FILE_NAME(RAW-FILE_NAME)=29-BT220720-P-29 3001.62 5xneg_BE4_01_34007_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3006.9	30-3006.9L	Sample source:Leaves | Genotype ID:3006 | Species:Fraxinus excelsior | Treatment:Susceptible European ash	RAW_FILE_NAME(RAW-FILE_NAME)=30-BT220720-L-30 3006.9 5x neg_GF4_01_33956_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3006.9	30-3006.9P	Sample source:Phloem | Genotype ID:3006 | Species:Fraxinus excelsior | Treatment:Susceptible European ash	RAW_FILE_NAME(RAW-FILE_NAME)=30-BT220720-P-30 3006.9 5x neg_BF4_01_34035_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3006.1	31-3006.1L	Sample source:Leaves | Genotype ID:3006 | Species:Fraxinus excelsior | Treatment:Susceptible European ash	RAW_FILE_NAME(RAW-FILE_NAME)=31-BT220720-L-31 3006.1 5x neg_GG4_01_33962_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3006.1	31-3006.1P	Sample source:Phloem | Genotype ID:3006 | Species:Fraxinus excelsior | Treatment:Susceptible European ash	RAW_FILE_NAME(RAW-FILE_NAME)=31-BT220720-P-31 3006.1 5x neg_BG4_01_34019_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3006.11	32-3006.11L	Sample source:Leaves | Genotype ID:3006 | Species:Fraxinus excelsior | Treatment:Susceptible European ash	RAW_FILE_NAME(RAW-FILE_NAME)=32-BT220720-L-32 3006.11 5xneg_GH4_01_33958_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3006.11	32-3006.11P	Sample source:Phloem | Genotype ID:3006 | Species:Fraxinus excelsior | Treatment:Susceptible European ash	RAW_FILE_NAME(RAW-FILE_NAME)=32-BT220720-P-32 3006.11 5xneg_BH4_01_34043_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3025.3	33-3025.3L	Sample source:Leaves | Genotype ID:3025 | Species:Fraxinus excelsior | Treatment:Susceptible European ash	RAW_FILE_NAME(RAW-FILE_NAME)=33-BT220720-L-33 3025.3 5x neg_GA5_01_33978_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3025.3	33-3025.3P	Sample source:Phloem | Genotype ID:3025 | Species:Fraxinus excelsior | Treatment:Susceptible European ash	RAW_FILE_NAME(RAW-FILE_NAME)=33-BT220720-P-33 3025.3 5x neg_BA5_01_34021_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3025.12	34-3025.12L	Sample source:Leaves | Genotype ID:3025 | Species:Fraxinus excelsior | Treatment:Susceptible European ash	RAW_FILE_NAME(RAW-FILE_NAME)=34-BT220720-L-34 3025.12 5xneg_GB5_01_33952_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3025.12	34-3025.12P	Sample source:Phloem | Genotype ID:3025 | Species:Fraxinus excelsior | Treatment:Susceptible European ash	RAW_FILE_NAME(RAW-FILE_NAME)=34-BT220720-P-34 3025.12 5xneg_BB5_01_34006_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3049.8	35-3049.8L	Sample source:Leaves | Genotype ID:3049 | Species:Fraxinus excelsior | Treatment:Susceptible European ash	RAW_FILE_NAME(RAW-FILE_NAME)=35-BT220720-L-35 3049.8 5x neg_GC5_01_33943_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3049.8	35-3049.8P	Sample source:Phloem | Genotype ID:3049 | Species:Fraxinus excelsior | Treatment:Susceptible European ash	RAW_FILE_NAME(RAW-FILE_NAME)=35-BT220720-P-35 3049.8 5x neg_BC5_01_34029_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3049.12	36-3049.12L	Sample source:Leaves | Genotype ID:3049 | Species:Fraxinus excelsior | Treatment:Susceptible European ash	RAW_FILE_NAME(RAW-FILE_NAME)=36-BT220720-L-36 3049.12 5xneg_GD5_01_33971_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3049.12	36-3049.12P	Sample source:Phloem | Genotype ID:3049 | Species:Fraxinus excelsior | Treatment:Susceptible European ash	RAW_FILE_NAME(RAW-FILE_NAME)=36-BT220720-P-36 3049.12 5xneg_BD5_01_33994_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3052.5	37-3052.5L	Sample source:Leaves | Genotype ID:3052 | Species:Fraxinus chinensis | Treatment:Tolerant Asian ash	RAW_FILE_NAME(RAW-FILE_NAME)=37-BT220720-L-37 3052.5 5x neg_GE5_01_33951_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3052.5	37-3052.5P	Sample source:Phloem | Genotype ID:3052 | Species:Fraxinus chinensis | Treatment:Tolerant Asian ash	RAW_FILE_NAME(RAW-FILE_NAME)=37-BT220720-P-37 3052.5 5x neg_BE5_01_34010_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3052.6	38-3052.6L	Sample source:Leaves | Genotype ID:3052 | Species:Fraxinus chinensis | Treatment:Tolerant Asian ash	RAW_FILE_NAME(RAW-FILE_NAME)=38-BT220720-L-38 3052.6 5x neg_GF5_01_33947_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3052.6	38-3052.6P	Sample source:Phloem | Genotype ID:3052 | Species:Fraxinus chinensis | Treatment:Tolerant Asian ash	RAW_FILE_NAME(RAW-FILE_NAME)=38-BT220720-P-38 3052.6 5x neg_BF5_01_34002_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3053.2	39-3053.2L	Sample source:Leaves | Genotype ID:3053 | Species:Fraxinus mandshurica | Treatment:Tolerant Asian ash	RAW_FILE_NAME(RAW-FILE_NAME)=39-BT220720-L-39 3053.2 5x neg_GG5_01_33940_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3053.2	39-3053.2P	Sample source:Phloem | Genotype ID:3053 | Species:Fraxinus mandshurica | Treatment:Tolerant Asian ash	RAW_FILE_NAME(RAW-FILE_NAME)=39-BT220720-P-39 3053.2 5x neg_BG5_01_34001_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3053.61	40-3053.61L	Sample source:Leaves | Genotype ID:3053 | Species:Fraxinus mandshurica | Treatment:Tolerant Asian ash	RAW_FILE_NAME(RAW-FILE_NAME)=40-BT220720-L-40 3053.61 5xneg_GH5_01_33968_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3053.61	40-3053.61P	Sample source:Phloem | Genotype ID:3053 | Species:Fraxinus mandshurica | Treatment:Tolerant Asian ash	RAW_FILE_NAME(RAW-FILE_NAME)=40-BT220720-P-40 3053.61 5xneg_BH5_01_34042_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3053.62	41-3053.62L	Sample source:Leaves | Genotype ID:3053 | Species:Fraxinus mandshurica | Treatment:Tolerant Asian ash	RAW_FILE_NAME(RAW-FILE_NAME)=41-BT220720-L-41 3053.62 5xneg_GA6_01_33949_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3053.62	41-3053.62P	Sample source:Phloem | Genotype ID:3053 | Species:Fraxinus mandshurica | Treatment:Tolerant Asian ash	RAW_FILE_NAME(RAW-FILE_NAME)=41-BT220720-P-41 3053.62 5xneg_BA6_01_33991_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3054.7	42-3054.7L	Sample source:Leaves | Genotype ID:3054 | Species:Fraxinus mandshurica | Treatment:Tolerant Asian ash	RAW_FILE_NAME(RAW-FILE_NAME)=42-BT220720-L-42 3054.7 5x neg_GB6_01_33941_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3054.7	42-3054.7P	Sample source:Phloem | Genotype ID:3054 | Species:Fraxinus mandshurica | Treatment:Tolerant Asian ash	RAW_FILE_NAME(RAW-FILE_NAME)=42-BT220720-P-42 3054.7 5x neg_BB6_01_34024_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3055.1	43-3055.1L	Sample source:Leaves | Genotype ID:3055 | Species:Fraxinus platypoda | Treatment:Tolerant Asian ash	RAW_FILE_NAME(RAW-FILE_NAME)=43-BT220720-L-43 3055.1 5x neg_GC6_01_33982_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3055.1	43-3055.1P	Sample source:Phloem | Genotype ID:3055 | Species:Fraxinus platypoda | Treatment:Tolerant Asian ash	RAW_FILE_NAME(RAW-FILE_NAME)=43-BT220720-P-43 3055.1 5x neg_BC6_01_34012_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3055.3	44-3055.3L	Sample source:Leaves | Genotype ID:3055 | Species:Fraxinus platypoda | Treatment:Tolerant Asian ash	RAW_FILE_NAME(RAW-FILE_NAME)=44-BT220720-L-44 3055.3 5x neg_GD6_01_33959_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3055.3	44-3055.3P	Sample source:Phloem | Genotype ID:3055 | Species:Fraxinus platypoda | Treatment:Tolerant Asian ash	RAW_FILE_NAME(RAW-FILE_NAME)=44-BT220720-P-44 3055.3 5x neg_BD6_01_34017_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3055.5	45-3055.5L	Sample source:Leaves | Genotype ID:3055 | Species:Fraxinus platypoda | Treatment:Tolerant Asian ash	RAW_FILE_NAME(RAW-FILE_NAME)=45-BT220720-L-45 3055.5 5x neg_GE6_01_33936_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3055.5	45-3055.5P	Sample source:Phloem | Genotype ID:3055 | Species:Fraxinus platypoda | Treatment:Tolerant Asian ash	RAW_FILE_NAME(RAW-FILE_NAME)=45-BT220720-P-45 3055.5 5x neg_BE6_01_34008_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3056.6	46-3056.6L	Sample source:Leaves | Genotype ID:3056 | Species:Fraxinus platypoda | Treatment:Tolerant Asian ash	RAW_FILE_NAME(RAW-FILE_NAME)=46-BT220720-L-46 3056.6 5x neg_GF6_01_33976_modified.mzML
SUBJECT_SAMPLE_FACTORS           	3056.6	46-3056.6P	Sample source:Phloem | Genotype ID:3056 | Species:Fraxinus platypoda | Treatment:Tolerant Asian ash	RAW_FILE_NAME(RAW-FILE_NAME)=46-BT220720-P-46 3056.6 5x neg_BF6_01_34028_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC mix 01	QC mix 01	Sample source:Mix of all samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC mix 01 neg_RA7_01_33922_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC mix 02	QC mix 02	Sample source:Mix of all samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC mix 02 neg_RA7_01_33923_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC mix 03	QC mix 03	Sample source:Mix of all samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC mix 03 neg_RA7_01_33930_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC mix 04	QC mix 04	Sample source:Mix of all samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC mix 04 neg_RA7_01_33931_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC mix 05	QC mix 05	Sample source:Mix of all samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC mix 05 neg_RA7_01_33932_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC mix 06	QC mix 06	Sample source:Mix of all samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC mix 06 neg_RA7_01_33939_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC mix 07	QC mix 07	Sample source:Mix of all samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC mix 07 neg_RA7_01_33946_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC mix 08	QC mix 08	Sample source:Mix of all samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC mix 08 neg_RA7_01_33953_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC mix 09	QC mix 09	Sample source:Mix of all samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC mix 09 neg_RA7_01_33960_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC mix 10	QC mix 10	Sample source:Mix of all samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC mix 10 neg_RA7_01_33967_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC mix 11	QC mix 11	Sample source:Mix of all samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC mix 11 neg_RA7_01_33974_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC mix 12	QC mix 12	Sample source:Mix of all samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC mix 12 neg_RA7_01_33981_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC mix 13	QC mix 13	Sample source:Mix of all samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC mix 13 neg_RA7_01_33990_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC mix 14	QC mix 14	Sample source:Mix of all samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC mix 14 neg_RA7_01_33997_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC mix 15	QC mix 15	Sample source:Mix of all samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC mix 15 neg_RA7_01_34004_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC mix 16	QC mix 16	Sample source:Mix of all samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC mix 16 neg_RA7_01_34011_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC mix 17	QC mix 17	Sample source:Mix of all samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC mix 17 neg_RA7_01_34018_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC mix 18	QC mix 18	Sample source:Mix of all samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC mix 18 neg_RA7_01_34025_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC mix 19	QC mix 19	Sample source:Mix of all samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC mix 19 neg_RA7_01_34032_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC mix 20	QC mix 20	Sample source:Mix of all samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC mix 20 neg_RA7_01_34039_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC mix 21	QC mix 21	Sample source:Mix of all samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC mix 21 neg_RA7_01_34044_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC leaves 01	QC leaves 01	Sample source:Mix of leaf samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC leaves 01 neg_RA5_01_33926_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC leaves 02	QC leaves 02	Sample source:Mix of leaf samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC leaves 02 neg_RA5_01_33927_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC leaves 03	QC leaves 03	Sample source:Mix of leaf samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC leaves 03 neg_RA5_01_33988_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC leaves 04	QC leaves 04	Sample source:Mix of leaf samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC leaves 04 neg_RA5_01_34045_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC leaves 05	QC leaves 05	Sample source:Mix of leaf samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC leaves 05 neg_RA5_01_34046_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC phloem 01	QC phloem 01	Sample source:Mix of phloem samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC phloem 01 neg_RA6_01_33928_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC phloem 02	QC phloem 02	Sample source:Mix of phloem samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC phloem 02 neg_RA6_01_33929_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC phloem 03	QC phloem 03	Sample source:Mix of phloem samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC phloem 03 neg_RA6_01_33989_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC phloem 04	QC phloem 04	Sample source:Mix of phloem samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC phloem 04 neg_RA6_01_34047_modified.mzML
SUBJECT_SAMPLE_FACTORS           	QC phloem 05	QC phloem 05	Sample source:Mix of phloem samples | Genotype ID:all | Species:all | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=QC phloem 05 neg_RA6_01_34048_modified.mzML
SUBJECT_SAMPLE_FACTORS           	Blank w IS 01	Blank w IS 01	Sample source:process Blank | Genotype ID:na | Species:na | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=Blank w IS 01 neg_RA4_01_33924_modified.mzML
SUBJECT_SAMPLE_FACTORS           	Blank w IS 02	Blank w IS 02	Sample source:process Blank | Genotype ID:na | Species:na | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=Blank w IS 02 neg_RA4_01_33925_modified.mzML
SUBJECT_SAMPLE_FACTORS           	Blank w IS 03	Blank w IS 03	Sample source:process Blank | Genotype ID:na | Species:na | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=Blank w IS 03 neg_RA4_01_34049_modified.mzML
SUBJECT_SAMPLE_FACTORS           	Blank w IS 04	Blank w IS 04	Sample source:process Blank | Genotype ID:na | Species:na | Treatment:-	RAW_FILE_NAME(RAW-FILE_NAME)=Blank w IS 04 neg_RA4_01_34050_modified.mzML
#COLLECTION
CO:COLLECTION_SUMMARY            	The material for this study was selected from a clonal field trial established
CO:COLLECTION_SUMMARY            	in 2016 in Snogeholm, Sjöbo, Sweden (55°32'57.4"N 13°42'29.4"E). The trial
CO:COLLECTION_SUMMARY            	consisted of 65 replicated ash genotypes: 56 tolerant F. excelsior trees
CO:COLLECTION_SUMMARY            	selected from around Sweden, four known susceptible F. excelsior genotypes, and
CO:COLLECTION_SUMMARY            	five genotypes representing three Fraxinus species of Asian origin. The Asian
CO:COLLECTION_SUMMARY            	ash include: F. mandshurica native to China, Japan, Korea, Eastern Russia; F.
CO:COLLECTION_SUMMARY            	platypoda native to China and Japan and, F. chinensis native to northern China,
CO:COLLECTION_SUMMARY            	Korea, Japan, south-east Russia (Wallander, 2013). Visually healthy leaf and
CO:COLLECTION_SUMMARY            	shoot samples (including bark and phloem) were collected in June 2020 from 13 F.
CO:COLLECTION_SUMMARY            	excelsior genotypes varying in disease severity to H. fraxineus (9 tolerant-ET
CO:COLLECTION_SUMMARY            	and 4 susceptible-ES) based on periodic assessments, and from all five resistant
CO:COLLECTION_SUMMARY            	Asian ash genotypes (AT) (Table S1). Up to five ramets (replicates) per genotype
CO:COLLECTION_SUMMARY            	were collected depending on the amount of available tissue. A total of 92
CO:COLLECTION_SUMMARY            	samples were collected in the field, 46 leaves and 46 phloem from the current
CO:COLLECTION_SUMMARY            	year shoots. Collected samples were placed in plastic bags and kept on dry ice
CO:COLLECTION_SUMMARY            	then stored at -80°C until further processing.
CO:COLLECTION_PROTOCOL_FILENAME  	Sampling024112.pdf
CO:SAMPLE_TYPE                   	Plant
CO:COLLECTION_LOCATION           	55°32'57.4"N 13°42'29.4"E
#TREATMENT
TR:TREATMENT_SUMMARY             	No treatment. This is a genotype study.
TR:TREATMENT_PROTOCOL_FILENAME   	Sampling024112.pdf
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Samples and tools were kept frozen in liquid nitrogen during all processing
SP:SAMPLEPREP_SUMMARY            	steps to minimize degradation or tissue oxidation. Bark including phloem was
SP:SAMPLEPREP_SUMMARY            	peeled from the twigs with a sterile scalpel. Bark and leaf tissue were ground
SP:SAMPLEPREP_SUMMARY            	separately in liquid nitrogen with porcelain mortars and pestles. For each
SP:SAMPLEPREP_SUMMARY            	sample, 100 mg of ground tissue was lyophilized for 24 hours. Metabolites
SP:SAMPLEPREP_SUMMARY            	extraction was performed by incubating the tissue with 500 μL of
SP:SAMPLEPREP_SUMMARY            	high-performance liquid chromatography grade MeOH containing schaftoside
SP:SAMPLEPREP_SUMMARY            	(Phytolab item no. 8332, Vestenbergsgreuth, Germany) as an internal standard at
SP:SAMPLEPREP_SUMMARY            	5 μM. Samples were centrifuged (4000 g, 2 min), and the supernatant was
SP:SAMPLEPREP_SUMMARY            	transferred to a new 2 mL microcentrifuge tube and stored at -20°C. The pellet
SP:SAMPLEPREP_SUMMARY            	was re-extracted as described above and the new supernatant was combined with
SP:SAMPLEPREP_SUMMARY            	previous extraction to get approximately 1000 μL of extract total per sample.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	Samples were 5-fold diluted with 75% MeOH in water and subjected to analysis by
CH:CHROMATOGRAPHY_SUMMARY        	ultra performance liquid chromatography coupled to tandem mass spectrometry
CH:CHROMATOGRAPHY_SUMMARY        	(UPLC- MS/MS). UPLC-MS/MS analysis was performed on a Dionex UltiMate 3000
CH:CHROMATOGRAPHY_SUMMARY        	Quaternary Rapid Separation UHPLC+ focused system (Thermo Fisher Scientific,
CH:CHROMATOGRAPHY_SUMMARY        	Germering, Germany. many). Separation was achieved on a Kinetex 1.7 μm XB-C18
CH:CHROMATOGRAPHY_SUMMARY        	column (100 × 2.1 mm, 1.7 μm, 100A).
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Thermo Dionex Ultimate 3000 RS
CH:COLUMN_NAME                   	Phenomenex Kinetex C18 (100 x 2.1mm,1.7um)
CH:SOLVENT_A                     	100% water; 0.05 % formic acid
CH:SOLVENT_B                     	100% Acetonitrile; 0.05 % formic acid
CH: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,
CH:FLOW_GRADIENT                 	50.0−53.0 min 60−100% B, 53.0−56.0 min 100% B, 56.0−56.5 min 100−3% B,
CH:FLOW_GRADIENT                 	and 56.5−60.0 min 3% B
CH:FLOW_RATE                     	300 µl/min
CH:COLUMN_TEMPERATURE            	30 °C
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
AN:OPERATOR_NAME                 	Christoph Crocoll
AN:DETECTOR_TYPE                 	ToF
AN:DATA_FORMAT                   	.d
#MS
MS:INSTRUMENT_NAME               	Bruker MicrOTOF II
MS:INSTRUMENT_TYPE               	QTOF
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	NEGATIVE
MS:MS_COMMENTS                   	The UHPLC was coupled to a Compact micrOTOF-Q mass spectrometer (Bruker, Bremen,
MS:MS_COMMENTS                   	Germany) equipped with an electrospray ion source (ESI) operated in either
MS:MS_COMMENTS                   	positive or negative ion mode. The ion spray voltage was maintained at -3900 V
MS:MS_COMMENTS                   	in negative ion mode. Dry temperature was set to 250 °C, and the dry gas flow
MS:MS_COMMENTS                   	was set to 8 L/min. Nitrogen was used as the dry gas, nebulizing gas, and
MS:MS_COMMENTS                   	collision gas. The nebulizing gas was set to 2.5 bar and collision energy to 15
MS:MS_COMMENTS                   	eV. MS spectra were acquired in an m/z range from 50 to 1000 m/z at a sampling
MS:MS_COMMENTS                   	rate of 3 Hz. MS/MS spectra were acquired in a range from 200-900 m/z.
MS:MS_COMMENTS                   	Na-formate clusters were used for mass calibration. MS raw data were processed
MS:MS_COMMENTS                   	with mzMine software version 2.53 (Pluskal et al., 2010) for peak detection,
MS:MS_COMMENTS                   	peak deconvolution, peak alignment and gap filling. The resulting peak-list were
MS:MS_COMMENTS                   	tentatively identified by matching values found in the KEGG COMPOUND Database
MS:MS_COMMENTS                   	https://www.genome.jp/kegg/compound/.
MS:CAPILLARY_VOLTAGE             	-3900 V
MS:COLLISION_ENERGY              	15 eV
MS:COLLISION_GAS                 	nitrogen
MS:DRY_GAS_FLOW                  	8 l/min
MS:DRY_GAS_TEMP                  	250 °C
MS:MS_RESULTS_FILE               	ST004232_AN007047_Results.txt	UNITS:Peak area	Has m/z:Yes	Has RT:Yes	RT units:Minutes
#END