#METABOLOMICS WORKBENCH Chanel5_20240704_035554 DATATRACK_ID:4976 STUDY_ID:ST003308 ANALYSIS_ID:AN005420 PROJECT_ID:PR002058
VERSION             	1
CREATED_ON             	July 5, 2024, 9:28 am
#PROJECT
PR:PROJECT_TITLE                 	An untargeted metabolic profiling strategy for the dissection of the oat (Avena
PR:PROJECT_TITLE                 	sativa L.) plant innate immune response to various Pseudomonas syringae
PR:PROJECT_TITLE                 	pathovars
PR:PROJECT_TYPE                  	Untargeted MS
PR:PROJECT_SUMMARY               	One of the most important characteristics that plants utilise to successfully
PR:PROJECT_SUMMARY               	defend themselves is the ability to rapidly identify potential threats in the
PR:PROJECT_SUMMARY               	surrounding environment. Plants rely on the perception of microbe-derived
PR:PROJECT_SUMMARY               	molecular pattern chemicals for this recognition, which initiates a number of
PR:PROJECT_SUMMARY               	induced defence reactions that ultimately increase plant resistance. The
PR:PROJECT_SUMMARY               	metabolome acts as a metabolic fingerprint of the biochemical activities that
PR:PROJECT_SUMMARY               	take place in a biological system under particular conditions and therefore
PR:PROJECT_SUMMARY               	provides a functional readout of the cellular mechanisms involved in a
PR:PROJECT_SUMMARY               	biological system. In this study, an untargeted metabolomics approach was
PR:PROJECT_SUMMARY               	applied to decipher the biochemical processes involved in oat plant defence
PR:PROJECT_SUMMARY               	responses to inoculation with various pathovars of Pseudomonas syringae
PR:PROJECT_SUMMARY               	(pathogenic and non-pathogenic on oat) such as P. syringae pv. coronafaciens
PR:PROJECT_SUMMARY               	(Ps-c), -pv. tabaci (Ps-t), -pv. tomato DC3000 (DC3000) and -pv. tomato DC3000
PR:PROJECT_SUMMARY               	hrcC mutant (hrcC−) and thereby identify signatory markers that are involved
PR:PROJECT_SUMMARY               	in host or nonhost defence responses. At the seedling growth stage, metabolic
PR:PROJECT_SUMMARY               	alterations in the Dunnart oat cultivar (tolerant to Ps-c) in response to
PR:PROJECT_SUMMARY               	inoculation with the respective Pseudomonas syringae pathovars were examined.
PR:PROJECT_SUMMARY               	Following inoculation, plants were monitored for symptom development and
PR:PROJECT_SUMMARY               	harvested at 2-, 4- and 6 d.p.i.. Methanolic metabolite extracts were prepared,
PR:PROJECT_SUMMARY               	and ultra-high-performance liquid chromatography (UHPLC) connected to a qTOF
PR:PROJECT_SUMMARY               	high-definition mass spectrometer was used to analyse the extracts. Chemometric
PR:PROJECT_SUMMARY               	modelling and multivariate statistical analysis revealed host- and time-related
PR:PROJECT_SUMMARY               	metabolic alterations that point to host and nonhost interactions in response to
PR:PROJECT_SUMMARY               	bacterial inoculation/infection. Metabolic profiles from further multivariate
PR:PROJECT_SUMMARY               	data analyses revealed a range of metabolite classes involved in the respective
PR:PROJECT_SUMMARY               	defence responses, including phenolic amides, saponins, phenolic acids,
PR:PROJECT_SUMMARY               	flavonoids, fatty acids, amino acids and alkaloids. The findings in this study
PR:PROJECT_SUMMARY               	allowed the elucidation of metabolic changes involved in oat defence responses
PR:PROJECT_SUMMARY               	to a range of pathovars of Pseudomonas syringae and ultimately contributed to a
PR:PROJECT_SUMMARY               	more comprehensive view of the oat plant metabolism under biotic stress during
PR:PROJECT_SUMMARY               	host vs nonhost interactions.
PR:INSTITUTE                     	University of Johannesburg
PR:DEPARTMENT                    	Biochemistry
PR:LABORATORY                    	Plant Metabolomics
PR:LAST_NAME                     	Pretorius
PR:FIRST_NAME                    	Chanel
PR:ADDRESS                       	Cnr Barry Hertzog and Napier Road, Richmond, Johannesburg
PR:EMAIL                         	chanelpretorius5@outlook.com
PR:PHONE                         	0660328667
#STUDY
ST:STUDY_TITLE                   	An untargeted metabolic profiling strategy for the dissection of the oat (Avena
ST:STUDY_TITLE                   	sativa L.) plant innate immune response to various Pseudomonas syringae
ST:STUDY_TITLE                   	pathovars
ST:STUDY_TYPE                    	Plant Metabolomics
ST:STUDY_SUMMARY                 	One of the most important characteristics that plants utilise to successfully
ST:STUDY_SUMMARY                 	defend themselves is the ability to rapidly identify potential threats in the
ST:STUDY_SUMMARY                 	surrounding environment. Plants rely on the perception of microbe-derived
ST:STUDY_SUMMARY                 	molecular pattern chemicals for this recognition, which initiates a number of
ST:STUDY_SUMMARY                 	induced defence reactions that ultimately increase plant resistance. The
ST:STUDY_SUMMARY                 	metabolome acts as a metabolic fingerprint of the biochemical activities that
ST:STUDY_SUMMARY                 	take place in a biological system under particular conditions and therefore
ST:STUDY_SUMMARY                 	provides a functional readout of the cellular mechanisms involved in a
ST:STUDY_SUMMARY                 	biological system. In this study, an untargeted metabolomics approach was
ST:STUDY_SUMMARY                 	applied to decipher the biochemical processes involved in oat plant defence
ST:STUDY_SUMMARY                 	responses to inoculation with various pathovars of Pseudomonas syringae
ST:STUDY_SUMMARY                 	(pathogenic and non-pathogenic on oat) such as P. syringae pv. coronafaciens
ST:STUDY_SUMMARY                 	(Ps-c), -pv. tabaci (Ps-t), -pv. tomato DC3000 (DC3000) and -pv. tomato DC3000
ST:STUDY_SUMMARY                 	hrcC mutant (hrcC−) and thereby identify signatory markers that are involved
ST:STUDY_SUMMARY                 	in host or nonhost defence responses. At the seedling growth stage, metabolic
ST:STUDY_SUMMARY                 	alterations in the Dunnart oat cultivar (tolerant to Ps-c) in response to
ST:STUDY_SUMMARY                 	inoculation with the respective Pseudomonas syringae pathovars were examined.
ST:STUDY_SUMMARY                 	Following inoculation, plants were monitored for symptom development and
ST:STUDY_SUMMARY                 	harvested at 2-, 4- and 6 d.p.i.. Methanolic metabolite extracts were prepared,
ST:STUDY_SUMMARY                 	and ultra-high-performance liquid chromatography (UHPLC) connected to a qTOF
ST:STUDY_SUMMARY                 	high-definition mass spectrometer was used to analyse the extracts. Chemometric
ST:STUDY_SUMMARY                 	modelling and multivariate statistical analysis revealed host- and time-related
ST:STUDY_SUMMARY                 	metabolic alterations that point to host and nonhost interactions in response to
ST:STUDY_SUMMARY                 	bacterial inoculation/infection. Metabolic profiles from further multivariate
ST:STUDY_SUMMARY                 	data analyses revealed a range of metabolite classes involved in the respective
ST:STUDY_SUMMARY                 	defence responses, including phenolic amides, saponins, phenolic acids,
ST:STUDY_SUMMARY                 	flavonoids, fatty acids, amino acids and alkaloids. The findings in this study
ST:STUDY_SUMMARY                 	allowed the elucidation of metabolic changes involved in oat defence responses
ST:STUDY_SUMMARY                 	to a range of pathovars of Pseudomonas syringae and ultimately contributed to a
ST:STUDY_SUMMARY                 	more comprehensive view of the oat plant metabolism under biotic stress during
ST:STUDY_SUMMARY                 	host vs nonhost interactions.
ST:INSTITUTE                     	University of Johannesburg
ST:DEPARTMENT                    	Biochemistry
ST:LABORATORY                    	Plant Metabolomics
ST:LAST_NAME                     	Pretorius
ST:FIRST_NAME                    	Chanel
ST:ADDRESS                       	Corner Kingsway and University Road, Auckland Park, Johannesburg, 2092
ST:EMAIL                         	chanelpretorius5@outlook.com
ST:PHONE                         	0660328667
#SUBJECT
SU:SUBJECT_TYPE                  	Plant
SU:SUBJECT_SPECIES               	Oat (Avena sativa L.)
#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           	-	DUNDC3000DAY2B1R1b	Sample source:Dunnart oat cultivar | Treatment:DC3000 | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNDC3000DAY2B1R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNDC3000DAY2B1R2b	Sample source:Dunnart oat cultivar | Treatment:DC3000 | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNDC3000DAY2B1R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNDC3000DAY2B1R3b	Sample source:Dunnart oat cultivar | Treatment:DC3000 | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNDC3000DAY2B1R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNDC3000DAY2B2R1b	Sample source:Dunnart oat cultivar | Treatment:DC3000 | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNDC3000DAY2B2R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNDC3000DAY2B2R2b	Sample source:Dunnart oat cultivar | Treatment:DC3000 | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNDC3000DAY2B2R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNDC3000DAY2B2R3b	Sample source:Dunnart oat cultivar | Treatment:DC3000 | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNDC3000DAY2B2R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNDC3000DAY2B3R1b	Sample source:Dunnart oat cultivar | Treatment:DC3000 | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNDC3000DAY2B3R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNDC3000DAY2B3R2b	Sample source:Dunnart oat cultivar | Treatment:DC3000 | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNDC3000DAY2B3R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNDC3000DAY2B3R3b	Sample source:Dunnart oat cultivar | Treatment:DC3000 | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNDC3000DAY2B3R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNDC3000DAY4B1R1b	Sample source:Dunnart oat cultivar | Treatment:DC3000 | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNDC3000DAY4B1R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNDC3000DAY4B1R2b	Sample source:Dunnart oat cultivar | Treatment:DC3000 | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNDC3000DAY4B1R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNDC3000DAY4B1R3b	Sample source:Dunnart oat cultivar | Treatment:DC3000 | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNDC3000DAY4B1R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNDC3000DAY4B2R1b	Sample source:Dunnart oat cultivar | Treatment:DC3000 | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNDC3000DAY4B2R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNDC3000DAY4B2R2b	Sample source:Dunnart oat cultivar | Treatment:DC3000 | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNDC3000DAY4B2R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNDC3000DAY4B2R3b	Sample source:Dunnart oat cultivar | Treatment:DC3000 | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNDC3000DAY4B2R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNDC3000DAY4B3R1b	Sample source:Dunnart oat cultivar | Treatment:DC3000 | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNDC3000DAY4B3R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNDC3000DAY4B3R2b	Sample source:Dunnart oat cultivar | Treatment:DC3000 | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNDC3000DAY4B3R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNDC3000DAY4B3R3b	Sample source:Dunnart oat cultivar | Treatment:DC3000 | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNDC3000DAY4B3R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNDC3000DAY6B1R1b	Sample source:Dunnart oat cultivar | Treatment:DC3000 | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNDC3000DAY6B1R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNDC3000DAY6B1R2b	Sample source:Dunnart oat cultivar | Treatment:DC3000 | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNDC3000DAY6B1R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNDC3000DAY6B1R3b	Sample source:Dunnart oat cultivar | Treatment:DC3000 | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNDC3000DAY6B1R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNDC3000DAY6B2R1b	Sample source:Dunnart oat cultivar | Treatment:DC3000 | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNDC3000DAY6B2R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNDC3000DAY6B2R2b	Sample source:Dunnart oat cultivar | Treatment:DC3000 | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNDC3000DAY6B2R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNDC3000DAY6B2R3b	Sample source:Dunnart oat cultivar | Treatment:DC3000 | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNDC3000DAY6B2R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNDC3000DAY6B3R1b	Sample source:Dunnart oat cultivar | Treatment:DC3000 | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNDC3000DAY6B3R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNDC3000DAY6B3R2b	Sample source:Dunnart oat cultivar | Treatment:DC3000 | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNDC3000DAY6B3R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNDC3000DAY6B3R3b	Sample source:Dunnart oat cultivar | Treatment:DC3000 | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNDC3000DAY6B3R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNHCDAY2B1R1b	Sample source:Dunnart oat cultivar | Treatment:Control | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNHCDAY2B1R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNHCDAY2B1R2b	Sample source:Dunnart oat cultivar | Treatment:Control | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNHCDAY2B1R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNHCDAY2B1R3b	Sample source:Dunnart oat cultivar | Treatment:Control | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNHCDAY2B1R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNHCDAY2B2R1b	Sample source:Dunnart oat cultivar | Treatment:Control | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNHCDAY2B2R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNHCDAY2B2R2b	Sample source:Dunnart oat cultivar | Treatment:Control | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNHCDAY2B2R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNHCDAY2B2R3b	Sample source:Dunnart oat cultivar | Treatment:Control | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNHCDAY2B2R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNHCDAY2B3R1b	Sample source:Dunnart oat cultivar | Treatment:Control | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNHCDAY2B3R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNHCDAY2B3R2b	Sample source:Dunnart oat cultivar | Treatment:Control | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNHCDAY2B3R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNHCDAY2B3R3b	Sample source:Dunnart oat cultivar | Treatment:Control | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNHCDAY2B3R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNHCDAY4B1R1b	Sample source:Dunnart oat cultivar | Treatment:Control | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNHCDAY4B1R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNHCDAY4B1R2b	Sample source:Dunnart oat cultivar | Treatment:Control | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNHCDAY4B1R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNHCDAY4B1R3b	Sample source:Dunnart oat cultivar | Treatment:Control | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNHCDAY4B1R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNHCDAY4B2R1b	Sample source:Dunnart oat cultivar | Treatment:Control | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNHCDAY4B2R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNHCDAY4B2R2b	Sample source:Dunnart oat cultivar | Treatment:Control | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNHCDAY4B2R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNHCDAY4B2R3b	Sample source:Dunnart oat cultivar | Treatment:Control | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNHCDAY4B2R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNHCDAY4B3R1b	Sample source:Dunnart oat cultivar | Treatment:Control | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNHCDAY4B3R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNHCDAY4B3R2b	Sample source:Dunnart oat cultivar | Treatment:Control | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNHCDAY4B3R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNHCDAY4B3R3b	Sample source:Dunnart oat cultivar | Treatment:Control | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNHCDAY4B3R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNHCDAY6B1R1b	Sample source:Dunnart oat cultivar | Treatment:Control | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNHCDAY6B1R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNHCDAY6B1R2b	Sample source:Dunnart oat cultivar | Treatment:Control | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNHCDAY6B1R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNHCDAY6B1R3b	Sample source:Dunnart oat cultivar | Treatment:Control | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNHCDAY6B1R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNHCDAY6B2R1b	Sample source:Dunnart oat cultivar | Treatment:Control | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNHCDAY6B2R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNHCDAY6B2R2b	Sample source:Dunnart oat cultivar | Treatment:Control | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNHCDAY6B2R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNHCDAY6B2R3b	Sample source:Dunnart oat cultivar | Treatment:Control | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNHCDAY6B2R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNHCDAY6B3R1b	Sample source:Dunnart oat cultivar | Treatment:Control | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNHCDAY6B3R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNHCDAY6B3R2b	Sample source:Dunnart oat cultivar | Treatment:Control | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNHCDAY6B3R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNHCDAY6B3R3b	Sample source:Dunnart oat cultivar | Treatment:Control | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNHCDAY6B3R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNHRCCDAY2B1R1b	Sample source:Dunnart oat cultivar | Treatment:HrcC mutant | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNHRCCDAY2B1R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNHRCCDAY2B1R2b	Sample source:Dunnart oat cultivar | Treatment:HrcC mutant | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNHRCCDAY2B1R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNHRCCDAY2B1R3b	Sample source:Dunnart oat cultivar | Treatment:HrcC mutant | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNHRCCDAY2B1R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNHRCCDAY2B2R1b	Sample source:Dunnart oat cultivar | Treatment:HrcC mutant | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNHRCCDAY2B2R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNHRCCDAY2B2R2b	Sample source:Dunnart oat cultivar | Treatment:HrcC mutant | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNHRCCDAY2B2R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNHRCCDAY2B2R3b	Sample source:Dunnart oat cultivar | Treatment:HrcC mutant | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNHRCCDAY2B2R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNHRCCDAY2B3R1b	Sample source:Dunnart oat cultivar | Treatment:HrcC mutant | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNHRCCDAY2B3R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNHRCCDAY2B3R2b	Sample source:Dunnart oat cultivar | Treatment:HrcC mutant | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNHRCCDAY2B3R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNHRCCDAY2B3R3b	Sample source:Dunnart oat cultivar | Treatment:HrcC mutant | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNHRCCDAY2B3R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNHRCCDAY4B1R1b	Sample source:Dunnart oat cultivar | Treatment:HrcC mutant | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNHRCCDAY4B1R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNHRCCDAY4B1R2b	Sample source:Dunnart oat cultivar | Treatment:HrcC mutant | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNHRCCDAY4B1R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNHRCCDAY4B1R3b	Sample source:Dunnart oat cultivar | Treatment:HrcC mutant | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNHRCCDAY4B1R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNHRCCDAY4B2R1b	Sample source:Dunnart oat cultivar | Treatment:HrcC mutant | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNHRCCDAY4B2R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNHRCCDAY4B2R2b	Sample source:Dunnart oat cultivar | Treatment:HrcC mutant | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNHRCCDAY4B2R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNHRCCDAY4B2R3b	Sample source:Dunnart oat cultivar | Treatment:HrcC mutant | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNHRCCDAY4B2R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNHRCCDAY4B3R1b	Sample source:Dunnart oat cultivar | Treatment:HrcC mutant | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNHRCCDAY4B3R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNHRCCDAY4B3R2b	Sample source:Dunnart oat cultivar | Treatment:HrcC mutant | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNHRCCDAY4B3R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNHRCCDAY4B3R3b	Sample source:Dunnart oat cultivar | Treatment:HrcC mutant | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNHRCCDAY4B3R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNHRCCDAY6B1R1b	Sample source:Dunnart oat cultivar | Treatment:HrcC mutant | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNHRCCDAY6B1R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNHRCCDAY6B1R2b	Sample source:Dunnart oat cultivar | Treatment:HrcC mutant | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNHRCCDAY6B1R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNHRCCDAY6B1R3b	Sample source:Dunnart oat cultivar | Treatment:HrcC mutant | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNHRCCDAY6B1R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNHRCCDAY6B2R1b	Sample source:Dunnart oat cultivar | Treatment:HrcC mutant | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNHRCCDAY6B2R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNHRCCDAY6B2R2b	Sample source:Dunnart oat cultivar | Treatment:HrcC mutant | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNHRCCDAY6B2R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNHRCCDAY6B2R3b	Sample source:Dunnart oat cultivar | Treatment:HrcC mutant | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNHRCCDAY6B2R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNHRCCDAY6B3R1b	Sample source:Dunnart oat cultivar | Treatment:HrcC mutant | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNHRCCDAY6B3R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNHRCCDAY6B3R2b	Sample source:Dunnart oat cultivar | Treatment:HrcC mutant | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNHRCCDAY6B3R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNHRCCDAY6B3R3b	Sample source:Dunnart oat cultivar | Treatment:HrcC mutant | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNHRCCDAY6B3R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSCDAY2B1R1b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas coronafaciens | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNPSCDAY2B1R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSCDAY2B1R2b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas coronafaciens | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNPSCDAY2B1R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSCDAY2B1R3b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas coronafaciens | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNPSCDAY2B1R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSCDAY2B2R1b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas coronafaciens | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNPSCDAY2B2R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSCDAY2B2R2b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas coronafaciens | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNPSCDAY2B2R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSCDAY2B2R3b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas coronafaciens | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNPSCDAY2B2R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSCDAY2B3R1b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas coronafaciens | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNPSCDAY2B3R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSCDAY2B3R2b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas coronafaciens | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNPSCDAY2B3R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSCDAY2B3R3b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas coronafaciens | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNPSCDAY2B3R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSCDAY4B1R1b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas coronafaciens | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNPSCDAY4B1R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSCDAY4B1R2b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas coronafaciens | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNPSCDAY4B1R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSCDAY4B1R3b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas coronafaciens | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNPSCDAY4B1R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSCDAY4B2R1b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas coronafaciens | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNPSCDAY4B2R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSCDAY4B2R2b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas coronafaciens | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNPSCDAY4B2R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSCDAY4B2R3b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas coronafaciens | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNPSCDAY4B2R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSCDAY4B3R1b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas coronafaciens | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNPSCDAY4B3R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSCDAY4B3R2b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas coronafaciens | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNPSCDAY4B3R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSCDAY4B3R3b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas coronafaciens | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNPSCDAY4B3R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSCDAY6B1R1b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas coronafaciens | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNPSCDAY6B1R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSCDAY6B1R2b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas coronafaciens | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNPSCDAY6B1R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSCDAY6B1R3b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas coronafaciens | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNPSCDAY6B1R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSCDAY6B2R1b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas coronafaciens | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNPSCDAY6B2R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSCDAY6B2R2b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas coronafaciens | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNPSCDAY6B2R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSCDAY6B2R3b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas coronafaciens | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNPSCDAY6B2R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSCDAY6B3R1b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas coronafaciens | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNPSCDAY6B3R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSCDAY6B3R2b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas coronafaciens | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNPSCDAY6B3R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSCDAY6B3R3b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas coronafaciens | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNPSCDAY6B3R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSTDAY2B1R1b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas tabaci | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNPSTDAY2B1R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSTDAY2B1R2b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas tabaci | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNPSTDAY2B1R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSTDAY2B1R3b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas tabaci | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNPSTDAY2B1R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSTDAY2B2R1b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas tabaci | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNPSTDAY2B2R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSTDAY2B2R2b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas tabaci | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNPSTDAY2B2R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSTDAY2B2R3b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas tabaci | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNPSTDAY2B2R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSTDAY2B3R1b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas tabaci | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNPSTDAY2B3R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSTDAY2B3R2b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas tabaci | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNPSTDAY2B3R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSTDAY2B3R3b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas tabaci | Days post inoculation:2 dpi	RAW_FILE_NAME(Raw file name)=DUNPSTDAY2B3R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSTDAY4B1R1b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas tabaci | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNPSTDAY4B1R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSTDAY4B1R2b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas tabaci | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNPSTDAY4B1R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSTDAY4B1R3b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas tabaci | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNPSTDAY4B1R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSTDAY4B2R1b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas tabaci | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNPSTDAY4B2R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSTDAY4B2R2b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas tabaci | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNPSTDAY4B2R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSTDAY4B2R3b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas tabaci | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNPSTDAY4B2R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSTDAY4B3R1b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas tabaci | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNPSTDAY4B3R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSTDAY4B3R2b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas tabaci | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNPSTDAY4B3R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSTDAY4B3R3b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas tabaci | Days post inoculation:4 dpi	RAW_FILE_NAME(Raw file name)=DUNPSTDAY4B3R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSTDAY6B1R1b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas tabaci | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNPSTDAY6B1R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSTDAY6B1R2b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas tabaci | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNPSTDAY6B1R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSTDAY6B1R3b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas tabaci | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNPSTDAY6B1R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSTDAY6B2R1b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas tabaci | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNPSTDAY6B2R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSTDAY6B2R2b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas tabaci | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNPSTDAY6B2R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSTDAY6B2R3b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas tabaci | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNPSTDAY6B2R3b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSTDAY6B3R1b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas tabaci | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNPSTDAY6B3R1b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSTDAY6B3R2b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas tabaci | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNPSTDAY6B3R2b
SUBJECT_SAMPLE_FACTORS           	-	DUNPSTDAY6B3R3b	Sample source:Dunnart oat cultivar | Treatment:Pseudomonas tabaci | Days post inoculation:6 dpi	RAW_FILE_NAME(Raw file name)=DUNPSTDAY6B3R3b
#COLLECTION
CO:COLLECTION_SUMMARY            	At the three-leaf growth stage the leaves were treated by spraying with the
CO:COLLECTION_SUMMARY            	Ps-c, Ps-t, DC3000 and hrcC− bacterial suspensions (prepared in PBS with 0.1 %
CO:COLLECTION_SUMMARY            	Tween 20), diluted to OD600 ≈0.3. The leaves were then harvested at 2, 4 and 6
CO:COLLECTION_SUMMARY            	days post inoculation (d.p.i.). The leaf material was quenched with liquid
CO:COLLECTION_SUMMARY            	nitrogen before being crushed into powder with a mortar and pestle. The samples
CO:COLLECTION_SUMMARY            	were weighed (1 g) suspended in 80% cold (4 °C) aqueous analytical grade
CO:COLLECTION_SUMMARY            	methanol at a 1:10 m/v ratio.
CO:SAMPLE_TYPE                   	Leaves
#TREATMENT
TR:TREATMENT_SUMMARY             	The leaves were treated by spraying with the Ps-c, Ps-t, DC3000 and hrcC−
TR:TREATMENT_SUMMARY             	bacterial suspensions (prepared in PBS with 0.1 % Tween 20), diluted to OD600
TR:TREATMENT_SUMMARY             	≈0.3. The vehicle control (VC) plants were sprayed with a solution free of the
TR:TREATMENT_SUMMARY             	bacteria and the healthy control (HC) groups were untreated (i.e., not sprayed
TR:TREATMENT_SUMMARY             	with either solution), all grown under normal growth conditions.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	The harvested leaf material was quenched with liquid nitrogen before being
SP:SAMPLEPREP_SUMMARY            	crushed into powder with a mortar and pestle. The samples were weighed (1 g)
SP:SAMPLEPREP_SUMMARY            	suspended in 80% cold (4 °C) aqueous analytical grade methanol (Romil's
SP:SAMPLEPREP_SUMMARY            	Chemistry, Cambridge, UK).at a 1:10 m/v ratio. The mixture was then homogenised
SP:SAMPLEPREP_SUMMARY            	with a probe sonicator (Bandelin Sonopuls, Berlin, Germany) at 55% power for 10
SP:SAMPLEPREP_SUMMARY            	seconds per sample. To avoid cross-contamination, equipment was cleansed between
SP:SAMPLEPREP_SUMMARY            	each sample. The homogenates were centrifuged at 5100 x g for 20 min at 4 °C in
SP:SAMPLEPREP_SUMMARY            	a benchtop centrifuge after which the supernatants were kept and concentrated by
SP:SAMPLEPREP_SUMMARY            	evaporating the methanol under vacuum to approximately 1 mL using a rotary
SP:SAMPLEPREP_SUMMARY            	evaporator set to 55 °C. The concentrated samples were transferred to 2 mL
SP:SAMPLEPREP_SUMMARY            	microcentrifuge tubes and dried in a centrifugal evaporator under vacuum. The
SP:SAMPLEPREP_SUMMARY            	dried extracts were then reconstituted by dissolving in 500 μL of 50% aqueous
SP:SAMPLEPREP_SUMMARY            	methanol (MilliQ deionised water and LC-grade methanol (Romil, Cambridge, UK).
SP:SAMPLEPREP_SUMMARY            	The samples were subsequently filtered through nylon syringe filters (0.22 μm)
SP:SAMPLEPREP_SUMMARY            	into chromatography vials fitted with 500 μL inserts, capped, and kept at 4 °C
SP:SAMPLEPREP_SUMMARY            	until analysis.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_SUMMARY        	Solvent A: MilliQ water containing 0.1% formic acid and 2.5% isopropanol;
CH:CHROMATOGRAPHY_SUMMARY        	Solvent B: Acetonitrile containing 0.1% formic acid and 2.5% isopropanol
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Waters Acquity
CH:COLUMN_NAME                   	Waters ACQUITY UPLC HSS T3 (150 x 2.1mm,1.7um)
CH:SOLVENT_A                     	97.5% MilliQ water/2.5% isopropanol ; 0.1% formic acid
CH:SOLVENT_B                     	97,5% acetonitrile/2.5% isopropanol; 0.1% formic acid
CH:FLOW_GRADIENT                 	Elution gradient via binary solvent system
CH:FLOW_RATE                     	0.4 mL/min
CH:COLUMN_TEMPERATURE            	60
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Waters Synapt G1
MS:INSTRUMENT_TYPE               	QTOF
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	NEGATIVE
MS:MS_COMMENTS                   	A high definition SYNAPT G1 quadrupole time-of-flight (qTOF) mass spectrometry
MS:MS_COMMENTS                   	system (Waters Corporation, Manchester, UK) was coupled to the UHPLC
MS:MS_COMMENTS                   	chromatography system to detect metabolites and acquire data in both positive
MS:MS_COMMENTS                   	and negative electrospray ionisation (ESI) operation modes. The controlling
MS:MS_COMMENTS                   	software was MassLynx XSTM (Waters, Manchester, UK). A reference calibrant,
MS:MS_COMMENTS                   	leucine encephalin (554.2615 Da) was used as the ‘lockmass’ calibrant and
MS:MS_COMMENTS                   	allowed for typical mass accuracies between 1 to 3 mDa. The respective capillary
MS:MS_COMMENTS                   	and sampling cone voltages were set as 2.5 kV and 30 V. The desolvation
MS:MS_COMMENTS                   	temperature used was 450 °C, with the source temperature set to 120 °C, cone
MS:MS_COMMENTS                   	gas flow was set to 50 L/h, and the desolvation gas flow set to 550 L/h. An m/z
MS:MS_COMMENTS                   	range of 50–1200 was set with a scan time of 0.1 s. The desolvation-,
MS:MS_COMMENTS                   	collision- and cone gas used at a flow rate of 700 L/h was high-purity nitrogen.
MS:MS_COMMENTS                   	Data was acquired using five different collision energies (MSE), ramping from
MS:MS_COMMENTS                   	0-50 eV to cause fragmentation of the initial ions to ensure that information
MS:MS_COMMENTS                   	regarding the fragmentation of the respective compounds could be obtained for
MS:MS_COMMENTS                   	downstream structural elucidation and metabolite annotation.
MS:MS_RESULTS_FILE               	ST003308_AN005420_Results.txt	UNITS:m/z	Has m/z:Yes	Has RT:Yes	RT units:Minutes
#END