#METABOLOMICS WORKBENCH annakisiala_20221212_112629 DATATRACK_ID:3641 STUDY_ID:ST002406 ANALYSIS_ID:AN003923 PROJECT_ID:PR001548
VERSION             	1
CREATED_ON             	December 12, 2022, 12:48 pm
#PROJECT
PR:PROJECT_TITLE                 	Silicon ameliorates clubroot responses in canola (Brassica napus): A
PR:PROJECT_TITLE                 	“multi-omics”-based investigation into possible mechanisms
PR:PROJECT_TYPE                  	MS quantitative analysis
PR:PROJECT_SUMMARY               	Clubroot disease, caused by Plasmodiophora brassicae Woronin results in severe
PR:PROJECT_SUMMARY               	yield losses in Brassica crops, including canola. Silicon (Si) mitigates several
PR:PROJECT_SUMMARY               	stresses and enhances plant resistance to phytopathogens. We investigated the
PR:PROJECT_SUMMARY               	effects of Si on clubroot disease symptoms in canola at two concentrations of Si
PR:PROJECT_SUMMARY               	(Si1.0 and Si0.5). In addition, the effects of Si on P. brassicae-induced gene
PR:PROJECT_SUMMARY               	expression, endogenous levels of phytohormones and metabolites were also
PR:PROJECT_SUMMARY               	studied. Si application reduced clubroot symptoms and improved plant growth
PR:PROJECT_SUMMARY               	under greenhouse conditions. Pathogen-induced transcript-level changes were
PR:PROJECT_SUMMARY               	affected by Si treatment to P. brassicae with genes related to antioxidant
PR:PROJECT_SUMMARY               	activity, phytohormone biosynthesis and signalling, nitrogen metabolism and
PR:PROJECT_SUMMARY               	secondary metabolism exhibiting differential expression. Endogenous levels of
PR:PROJECT_SUMMARY               	several phytohormones (e.g., auxin, cytokinin, salicylic acid and abscisic
PR:PROJECT_SUMMARY               	acid), amino acids and secondary metabolites (e.g., glucosinolates) were
PR:PROJECT_SUMMARY               	affected by Si. This is the first report that Si ameliorates clubroot symptoms
PR:PROJECT_SUMMARY               	and its possible mode of action.
PR:INSTITUTE                     	Trent University
PR:DEPARTMENT                    	Biology
PR:LABORATORY                    	Emery Lab
PR:LAST_NAME                     	Kisiala
PR:FIRST_NAME                    	Anna
PR:ADDRESS                       	1600 West Bank Drive, Trent University, Peterborough, ON, K9L 0G2, Canada
PR:EMAIL                         	annakisiala@trentu.ca
PR:PHONE                         	7057481011
PR:FUNDING_SOURCE                	Alberta Agriculture and Forestry
PR:CONTRIBUTORS                  	Ananya Sarkar, Anna Kisiala, Dinesh Adhikary, Urmila Basu, Neil Emery, Rahman,
PR:CONTRIBUTORS                  	Nat N. V. Kav
#STUDY
ST:STUDY_TITLE                   	Silicon ameliorates clubroot responses in canola (Brassica napus): A
ST:STUDY_TITLE                   	“multi-omics”-based investigation into possible mechanisms
ST:STUDY_TYPE                    	Timecourse experiment
ST:STUDY_SUMMARY                 	Clubroot disease, caused by Plasmodiophora brassicae Woronin results in severe
ST:STUDY_SUMMARY                 	yield losses in Brassica crops, including canola. Silicon (Si) mitigates several
ST:STUDY_SUMMARY                 	stresses and enhances plant resistance to phytopathogens. We investigated the
ST:STUDY_SUMMARY                 	effects of Si on clubroot disease symptoms in canola at two concentrations of Si
ST:STUDY_SUMMARY                 	(Si1.0 and Si0.5). In addition, the effects of Si on P. brassicae-induced gene
ST:STUDY_SUMMARY                 	expression, endogenous levels of phytohormones and metabolites were also
ST:STUDY_SUMMARY                 	studied. Si application reduced clubroot symptoms and improved plant growth
ST:STUDY_SUMMARY                 	under greenhouse conditions. Pathogen-induced transcript-level changes were
ST:STUDY_SUMMARY                 	affected by Si treatment to P. brassicae with genes related to antioxidant
ST:STUDY_SUMMARY                 	activity, phytohormone biosynthesis and signalling, nitrogen metabolism and
ST:STUDY_SUMMARY                 	secondary metabolism exhibiting differential expression. Endogenous levels of
ST:STUDY_SUMMARY                 	several phytohormones (e.g., auxin, cytokinin, salicylic acid and abscisic
ST:STUDY_SUMMARY                 	acid), amino acids and secondary metabolites (e.g., glucosinolates) were
ST:STUDY_SUMMARY                 	affected by Si. This is the first report that Si ameliorates clubroot symptoms
ST:STUDY_SUMMARY                 	and its possible mode of action.
ST:INSTITUTE                     	Trent University
ST:DEPARTMENT                    	Biology
ST:LABORATORY                    	Emery Lab
ST:LAST_NAME                     	Kisiala
ST:FIRST_NAME                    	Anna
ST:ADDRESS                       	1600 West Bank Drive, Trent University, Peterborough, ON, K9L 0G2, Canada
ST:EMAIL                         	annakisiala@trentu.ca
ST:PHONE                         	7057481011
#SUBJECT
SU:SUBJECT_TYPE                  	Plant
SU:SUBJECT_SPECIES               	Brassica napus
SU:TAXONOMY_ID                   	3708
#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           	-	C18-1-pos	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C1-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-1-neg	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C1-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-1-pos	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H1-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-1-neg	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H1-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-2-pos	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C2-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-2-neg	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C2-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-2-pos	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H2-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-2-neg	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H2-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-3-pos	Developmental stage (DPI):day 7 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C3-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-3-neg	Developmental stage (DPI):day 7 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C3-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-3-pos	Developmental stage (DPI):day 7 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H3-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-3-neg	Developmental stage (DPI):day 7 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H3-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-4-pos	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C4-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-4-neg	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C4-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-4-pos	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H4-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-4-neg	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H4-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-5-pos	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C5-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-5-neg	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C5-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-5-pos	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H5-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-5-neg	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H5-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-6-pos	Developmental stage (DPI):day 7 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C6-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-6-neg	Developmental stage (DPI):day 7 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C6-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-6-pos	Developmental stage (DPI):day 7 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H6-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-6-neg	Developmental stage (DPI):day 7 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H6-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-7-pos	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C7-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-7-neg	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C7-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-7-pos	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H7-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-7-neg	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H7-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-8-pos	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C8-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-8-neg	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C8-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-8-pos	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H8-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-8-neg	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H8-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-9-pos	Developmental stage (DPI):day 7 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C9-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-9-neg	Developmental stage (DPI):day 7 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C9-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-9-pos	Developmental stage (DPI):day 14 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H9-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-9-neg	Developmental stage (DPI):day 14 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H9-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-10-pos	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C10-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-10-neg	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C10-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-10-pos	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H10-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-10-neg	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H10-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-11-pos	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C11-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-11-neg	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C11-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-11-pos	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H11-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-11-neg	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H11-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-12-pos	Developmental stage (DPI):day 14 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C12-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-12-neg	Developmental stage (DPI):day 14 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C12-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-12-pos	Developmental stage (DPI):day 14 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H12-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-12-neg	Developmental stage (DPI):day 14 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H12-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-13-pos	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C13-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-13-neg	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C13-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-13-pos	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H13-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-13-neg	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H13-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-14-pos	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C14-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-14-neg	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C14-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-14-pos	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H14-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-14-neg	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H14-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-15-pos	Developmental stage (DPI):day 14 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C15-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-15-neg	Developmental stage (DPI):day 14 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C15-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-15-pos	Developmental stage (DPI):day 14 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H15-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-15-neg	Developmental stage (DPI):day 14 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H15-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-16-pos	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C16-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-16-neg	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C16-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-16-pos	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H16-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-16-neg	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H16-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-17-pos	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C17-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-17-neg	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C17-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-17-pos	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H17-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-17-neg	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H17-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-18-pos	Developmental stage (DPI):day 14 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C18-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-18-neg	Developmental stage (DPI):day 14 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C18-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-18-pos	Developmental stage (DPI):day 14 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H18-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-18-neg	Developmental stage (DPI):day 14 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H18-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-19-pos	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C19-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-19-neg	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C19-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-19-pos	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H19-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-19-neg	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H19-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-20-pos	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C20-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-20-neg	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C20-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-20-pos	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H20-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-20-neg	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H20-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-21-pos	Developmental stage (DPI):day 14 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C21-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-21-neg	Developmental stage (DPI):day 14 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C21-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-21-pos	Developmental stage (DPI):day 14 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H21-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-21-neg	Developmental stage (DPI):day 14 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H21-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-22-pos	Developmental stage (DPI):day 21 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C22-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-22-neg	Developmental stage (DPI):day 21 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C22-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-22-pos	Developmental stage (DPI):day 21 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H22-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-22-neg	Developmental stage (DPI):day 21 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H22-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-23-pos	Developmental stage (DPI):day 21 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C23-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-23-neg	Developmental stage (DPI):day 21 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C23-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-23-pos	Developmental stage (DPI):day 21 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H23-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-23-neg	Developmental stage (DPI):day 21 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H23-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-24-pos	Developmental stage (DPI):day 21 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C24-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-24-neg	Developmental stage (DPI):day 21 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C24-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-24-pos	Developmental stage (DPI):day 21 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H24-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-24-neg	Developmental stage (DPI):day 21 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H24-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-25-pos	Developmental stage (DPI):day 21 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C25-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-25-neg	Developmental stage (DPI):day 21 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C25-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-25-pos	Developmental stage (DPI):day 21 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H25-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-25-neg	Developmental stage (DPI):day 21 | Silicon treatment [g]:0.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H25-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-26-pos	Developmental stage (DPI):day 21 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C26-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-26-neg	Developmental stage (DPI):day 21 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C26-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-26-pos	Developmental stage (DPI):day 21 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H26-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-26-neg	Developmental stage (DPI):day 21 | Silicon treatment [g]:0.5	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H26-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-27-pos	Developmental stage (DPI):day 21 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C27-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	C18-27-neg	Developmental stage (DPI):day 21 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=C27-neg.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-27-pos	Developmental stage (DPI):day 21 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H27-pos.mzXML
SUBJECT_SAMPLE_FACTORS           	-	HILIC-27-neg	Developmental stage (DPI):day 21 | Silicon treatment [g]:1.0	Organism=Brassica napus; Cultivar=Canola (HiQ); Phenotype=susceptible to Plasmodiophora brassicae (P3H); Tissue=roots; RAW_FILE_NAME=H27-neg.mzXML
#COLLECTION
CO:COLLECTION_SUMMARY            	A total of 27 samples from the three treatments (PC, Si0.5 and Si1.0) were used
CO:COLLECTION_SUMMARY            	for metabolomic analyses at 7-, 14- and 21 dpi. Hi-Q seedlings were grown in the
CO:COLLECTION_SUMMARY            	soil-mix as previously described with/without Si amendment, inoculated with P.
CO:COLLECTION_SUMMARY            	brassicae, and subsequently, washed thoroughly, flash-frozen in liquid nitrogen
CO:COLLECTION_SUMMARY            	and stored at -80°C until analysis. Each biological replicate was comprised of
CO:COLLECTION_SUMMARY            	5 individual plants per treatment, and three biological replicates were
CO:COLLECTION_SUMMARY            	generated from three independent experiments. A total of 27 samples were
CO:COLLECTION_SUMMARY            	generated for transcriptomics and metabolomics analyses [3 treatments (PC, Si0.5
CO:COLLECTION_SUMMARY            	and Si1.0) x 3 time-points (7-, 14- and 21 dpi) x 3 biological replicates].
CO:COLLECTION_PROTOCOL_FILENAME  	Collection_protocol
CO:SAMPLE_TYPE                   	Plant
CO:COLLECTION_LOCATION           	Alberta, Canada
CO:COLLECTION_FREQUENCY          	7, 14 21 DPI
CO:VOLUMEORAMOUNT_COLLECTED      	0.07g per sample
CO:STORAGE_CONDITIONS            	-80℃
CO:COLLECTION_VIALS              	2 mL round-bottom Eppendorf tubes
CO:STORAGE_VIALS                 	2 mL round-bottom Eppendorf tubes
CO:COLLECTION_TUBE_TEMP          	-20
#TREATMENT
TR:TREATMENT_SUMMARY             	Sodium silicate (Na2SiO3, Mol. Wt: 122.06 g/L), obtained from Sigma-Aldrich (St.
TR:TREATMENT_SUMMARY             	Louis, US), was mixed with 100 g Sunshine Professional Growing Mix (Sun Gro
TR:TREATMENT_SUMMARY             	Horticulture, Seba Beach, Canada) and the following mixtures (treatments) were
TR:TREATMENT_SUMMARY             	produced: 0.0 g Si (control), 0.1 g Si (Si:soil in 1:1000, wt/wt), 0.25 g
TR:TREATMENT_SUMMARY             	(Si:soil in 1:400, wt/wt), 0.5 g (Si:soil in 1:200, wt/wt), 0.75 g (Si:soil in
TR:TREATMENT_SUMMARY             	3:400, wt/wt) and 1.0 g (Si:soil in 1:100, wt/wt). These treatments, hereafter,
TR:TREATMENT_SUMMARY             	reported as Si0.0 or PC, Si0.1, Si0.25, Si0.5, Si0.75 and Si1.0, respectively.
TR:TREATMENT_SUMMARY             	After mixing thoroughly, the pH of the soil mix was determined prior to seeding.
TR:TREATMENT_SUMMARY             	A clubroot susceptible spring canola (Brassica napus L.) cultivar Hi-Q,
TR:TREATMENT_SUMMARY             	developed at the University of Alberta, was used in this study. The plants were
TR:TREATMENT_SUMMARY             	grown under greenhouse conditions (22/19°C and 16/8h photoperiod) on the
TR:TREATMENT_SUMMARY             	above-mentioned six soil media (treatments) containing varying concentration of
TR:TREATMENT_SUMMARY             	Si. A total of 30 plants, from three replications of each treatment (i.e. 10
TR:TREATMENT_SUMMARY             	plants per replication) were used. A single spore suspension of P. brassicae
TR:TREATMENT_SUMMARY             	pathotype 3, classified as pathotype 3H (P3H) under the Canadian Clubroot
TR:TREATMENT_SUMMARY             	Differential (CCD) set, was prepared by homogenizing the clubroot galls, and
TR:TREATMENT_SUMMARY             	used to inoculate the seedlings following previously described procedures.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	A total of 27 samples from the three treatments (PC, Si0.5 and Si1.0) were used
SP:SAMPLEPREP_SUMMARY            	for metabolomic analyses at 7-, 14- and 21 dpi. Hi-Q seedlings were grown in the
SP:SAMPLEPREP_SUMMARY            	soil-mix as previously described with/without Si amendment, inoculated with P.
SP:SAMPLEPREP_SUMMARY            	brassicae, and subsequently, washed thoroughly, flash-frozen in liquid nitrogen
SP:SAMPLEPREP_SUMMARY            	and stored at -80°C until analysis. Each biological replicate was comprised of
SP:SAMPLEPREP_SUMMARY            	5 individual plants per treatment, and three biological replicates were
SP:SAMPLEPREP_SUMMARY            	generated from three independent experiments. A total of 27 samples were
SP:SAMPLEPREP_SUMMARY            	generated for transcriptomics and metabolomics analyses [3 treatments (PC, Si0.5
SP:SAMPLEPREP_SUMMARY            	and Si1.0) x 3 time-points (7-, 14- and 21 dpi) x 3 biological replicates].
SP:SAMPLEPREP_PROTOCOL_FILENAME  	Sample Prep protocol
SP:PROCESSING_STORAGE_CONDITIONS 	-20℃
SP:EXTRACTION_METHOD             	50% ACN
SP:EXTRACT_CLEANUP               	HLB SPE
SP:EXTRACT_STORAGE               	-20℃
SP:SAMPLE_RESUSPENSION           	Primary metabolites - 90% ACN, glucosinolates - 5% ACN
SP:SAMPLE_SPIKING                	Primary metabolites - Labeled amino acid mix; Glucosinolates - labeled CK mix
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Thermo Dionex Ultimate 3000
CH:COLUMN_NAME                   	Phenomonex Kinetex C18 (50 x 2.1mm, 2.6um)
CH:SOLVENT_A                     	100% water; 0.08% acetic acid
CH:SOLVENT_B                     	100% acetonitrile; 0.08% acetic acid
CH:FLOW_GRADIENT                 	Mobile phase 100% A held for 1.25 min, then decreased to 50% over 1.75 min and
CH:FLOW_GRADIENT                 	to 0% over the next 0.5 min, held at 0% for 2 min and returned to 100% over 0.5
CH:FLOW_GRADIENT                 	min for 4 min of column re-equilibration.
CH:FLOW_RATE                     	0.3 ml/min
CH:COLUMN_TEMPERATURE            	RT
CH:INJECTION_TEMPERATURE         	4
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Thermo Q Exactive Orbitrap
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	NEGATIVE
MS:MS_COMMENTS                   	Processing of the full scan and ddMS2 data was conducted using the Xcalibur 4.1
MS:MS_COMMENTS                   	software (Thermo Scientific, Waltham, US). Metabolites were identified by
MS:MS_COMMENTS                   	accurate mass, comparison of retention times to authentic standards or by
MS:MS_COMMENTS                   	accurate mass and comparison of fragmentation patterns to MS/MS databases
MS:MS_COMMENTS                   	(METLIN, PubChem).
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS	normalized relative level
MS_METABOLITE_DATA_START
Samples	C18-1-neg	C18-2-neg	C18-3-neg	C18-4-neg	C18-5-neg	C18-6-neg	C18-7-neg	C18-8-neg	C18-9-neg	C18-10-neg	C18-11-neg	C18-12-neg	C18-13-neg	C18-14-neg	C18-15-neg	C18-16-neg	C18-17-neg	C18-18-neg	C18-19-neg	C18-20-neg	C18-21-neg	C18-22-neg	C18-23-neg	C18-24-neg	C18-25-neg	C18-26-neg	C18-27-neg
Factors	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.0	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.5	Developmental stage (DPI):day 7 | Silicon treatment [g]:1.0	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.0	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.5	Developmental stage (DPI):day 7 | Silicon treatment [g]:1.0	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.0	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.5	Developmental stage (DPI):day 7 | Silicon treatment [g]:1.0	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.0	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.5	Developmental stage (DPI):day 14 | Silicon treatment [g]:1.0	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.0	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.5	Developmental stage (DPI):day 14 | Silicon treatment [g]:1.0	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.0	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.5	Developmental stage (DPI):day 14 | Silicon treatment [g]:1.0	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.0	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.5	Developmental stage (DPI):day 14 | Silicon treatment [g]:1.0	Developmental stage (DPI):day 21 | Silicon treatment [g]:0.0	Developmental stage (DPI):day 21 | Silicon treatment [g]:0.5	Developmental stage (DPI):day 21 | Silicon treatment [g]:1.0	Developmental stage (DPI):day 21 | Silicon treatment [g]:0.0	Developmental stage (DPI):day 21 | Silicon treatment [g]:0.5	Developmental stage (DPI):day 21 | Silicon treatment [g]:1.0
1,1'-diindolylmethane	1.019	0.948	1.002	1.413	1.102	1.006	0.620	0.991	0.901	0.798	0.748	0.535	0.891	1.003	0.985	0.982	0.661	0.833	0.760	0.716	0.696	0.880	0.279	0.054	1.106	0.919	1.025
4-methoxyglucobrassicin	0.244	0.045	0.054	0.089	0.406	0.186	0.334	0.210	0.066	0.452	1.351	0.763	0.248	0.222	0.124	0.049	0.042	0.066	1.324	1.223	0.544	0.128	0.128	0.151	0.081	0.228	0.108
Brassicanal A	0.052	0.091	0.026	0.273	0.034	0.370	0.210	0.130	0.037	1.332	0.863	0.947	0.150	0.045	0.080	0.055	0.050	0.080	0.601	0.952	0.685	0.194	0.244	0.183	0.048	0.010	0.015
Brassicanate A	0.505	0.504	0.541	0.587	0.006	0.820	0.621	0.260	0.514	0.678	1.129	0.970	1.394	0.477	0.537	0.244	0.356	0.508	0.320	0.211	0.233	0.575	0.516	0.540	0.542	0.448	0.435
Glucobrassicin	0.137	0.051	0.089	0.083	0.211	0.178	0.118	0.095	0.062	0.289	1.351	0.736	0.183	0.093	0.041	0.043	0.036	0.058	0.314	0.421	0.156	0.062	0.064	0.064	0.089	0.265	0.064
Gluconasturtiin	0.078	0.041	0.070	0.066	0.265	0.139	0.207	0.188	0.097	0.163	0.767	0.370	0.416	0.196	0.114	0.058	0.055	0.290	0.047	0.067	0.044	0.105	0.031	0.106	0.256	1.278	0.446
Indole-3-carboxaldehyde	0.343	0.411	0.299	0.414	0.621	0.561	0.573	0.466	0.332	0.675	0.593	0.632	0.413	0.270	0.296	0.287	0.270	0.260	1.162	1.340	0.868	0.400	0.572	0.461	0.339	0.171	0.185
Neoglucobrassicin	0.402	0.110	0.194	0.141	0.516	0.373	0.446	0.272	0.169	0.416	1.351	0.737	0.584	0.159	0.161	0.095	0.112	0.239	0.172	0.226	0.155	0.157	0.062	0.119	0.236	0.384	0.209
Rutalexin	0.000	0.002	0.001	0.021	0.010	0.003	0.025	0.022	0.004	0.183	0.193	0.647	0.020	0.067	0.006	0.002	0.003	0.005	0.984	1.340	1.023	0.027	0.051	0.089	0.004	0.013	0.169
Syringin	0.066	0.090	0.072	0.103	0.105	0.110	0.098	0.293	0.046	0.379	0.587	0.716	0.778	0.490	0.514	0.159	0.339	0.359	1.535	0.974	1.324	0.623	0.520	0.785	0.274	0.242	0.189
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	quanitfied m/z	PubChemID	KEGG
1,1'-diindolylmethane	245.10841
4-methoxyglucobrassicin	477.06430	656563	C08423
Brassicanal A	190.03320	443054 	C11048  
Brassicanate A	220.04377	11481434
Glucobrassicin	447.05374	6602378	C05837
Gluconasturtiin	422.05849	9548618	C08417
Indole-3-carboxaldehyde	144.04548	10256	C08493 
Neoglucobrassicin	477.06430	5485217	C08424
Rutalexin	231.02336	11195626
Syringin	371.13476	5316860	C01533  
METABOLITES_END
#END