#METABOLOMICS WORKBENCH annakisiala_20221212_112629 DATATRACK_ID:3641 STUDY_ID:ST002406 ANALYSIS_ID:AN003921
VERSION                          	1
CREATED_ON                       	12-16-2022
#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:DOI                           	http://dx.doi.org/10.21228/M8M42M
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
ST:SUBMIT_DATE                   	2022-12-12
#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]Additional sample data
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           	-	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           	-	C18-13-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           	-	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           	-	C18-16-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           	-	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           	-	C18-19-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           	-	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           	-	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           	-	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-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           	-	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-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           	-	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-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           	-	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           	-	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           	-	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           	-	C18-14-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           	-	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           	-	C18-17-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           	-	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           	-	C18-20-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           	-	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           	-	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           	-	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-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           	-	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-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           	-	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-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           	-	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           	-	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           	-	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           	-	C18-15-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           	-	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           	-	C18-18-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           	-	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           	-	C18-21-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           	-	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           	-	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           	-	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-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           	-	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-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           	-	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-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           	-	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-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           	-	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           	-	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           	-	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           	-	C18-25-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           	-	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           	-	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           	-	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-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           	-	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           	-	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           	-	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           	-	C18-26-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           	-	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           	-	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           	-	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-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           	-	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           	-	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           	-	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           	-	C18-27-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           	-	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           	-	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           	-	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-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
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           	-	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           	-	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           	-	C18-4-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           	-	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           	-	C18-7-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           	-	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           	-	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           	-	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-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           	-	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-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           	-	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           	-	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           	-	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           	-	C18-5-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           	-	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           	-	C18-8-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           	-	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           	-	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           	-	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-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           	-	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-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           	-	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           	-	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           	-	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           	-	C18-6-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           	-	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           	-	C18-9-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           	-	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           	-	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           	-	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-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           	-	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
#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:INSTRUMENT_NAME               	Thermo Dionex Ultimate 3000
CH:COLUMN_NAME                   	Agilent InfinityLab Poroshell 120 HILIC-Z (100 x 2.1mm, 2.7um)
CH:COLUMN_TEMPERATURE            	RT
CH:FLOW_GRADIENT                 	Mobile phase 100% B decreased to 90% over 2.5 min and to 50% over the next 5 min
CH:FLOW_GRADIENT                 	and returned to 100% over 0.5 min for 12 min of column re-equilibration.
CH:FLOW_RATE                     	0.2 ml/min
CH:INJECTION_TEMPERATURE         	4
CH:SOLVENT_A                     	100% water; 10 mM ammonium bicarbonate
CH:SOLVENT_B                     	95% acetonitrile; 10 mM ammonium bicarbonate
CH:CHROMATOGRAPHY_TYPE           	HILIC
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Thermo Q Exactive Orbitrap
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	ESI
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:ION_MODE                      	NEGATIVE
#MS_METABOLITE_DATA
MS_METABOLITE_DATA:UNITS         	normalized relative level
MS_METABOLITE_DATA_START
Samples	HILIC-10-neg	HILIC-13-neg	HILIC-16-neg	HILIC-19-neg	HILIC-11-neg	HILIC-14-neg	HILIC-17-neg	HILIC-20-neg	HILIC-12-neg	HILIC-15-neg	HILIC-18-neg	HILIC-21-neg	HILIC-9-neg	HILIC-22-neg	HILIC-25-neg	HILIC-23-neg	HILIC-26-neg	HILIC-24-neg	HILIC-27-neg	HILIC-1-neg	HILIC-4-neg	HILIC-7-neg	HILIC-2-neg	HILIC-5-neg	HILIC-8-neg	HILIC-3-neg	HILIC-6-neg
Factors	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.0	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.0	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.0	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]:0.5	Developmental stage (DPI):day 14 | Silicon treatment [g]:0.5	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]:1.0	Developmental stage (DPI):day 14 | Silicon treatment [g]:1.0	Developmental stage (DPI):day 14 | Silicon treatment [g]:1.0	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]:0.0	Developmental stage (DPI):day 21 | Silicon treatment [g]:0.5	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]:1.0	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.0	Developmental stage (DPI):day 7 | Silicon treatment [g]:0.0	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]:0.5	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]:1.0	
Asparagine	0.1380	0.0500	0.0690	0.2740	0.2470	0.0720	0.0960	0.5880	0.1390	0.1200	0.1360	0.3250	0.0990	1.9600	0.5790	1.5980	0.5880	3.2030	0.3630	0.3270	0.0320	0.0520	0.2460	0.0800	0.0410	0.0590	0.0550
Aspartate	0.4950	0.5150	0.3430	0.5070	0.2270	0.3610	0.2490	0.3540	0.0370	0.3090	0.2590	0.4010	0.2890	1.2530	1.0540	0.9810	1.5280	1.2940	0.7370	0.4400	0.1200	0.1580	0.3160	0.1020	0.0220	0.0370	
Coniferin	1.3850	1.0810	0.3190	1.1900	0.7150	1.0270	0.2550	1.7210	4.8510	0.3490	0.3440	4.1380	0.8750	2.2070	1.1460	0.8100	1.4250	1.7080	1.5100	0.1640	2.1530	1.1310	0.3870	2.0320	3.2380	3.0910	1.9250
Leucine	0.8350	0.9100	0.2360	2.2820	0.8480	0.6600	0.3570	2.7870	2.5860	0.4180	0.2570	3.3990	1.3950	1.3860	0.9250	1.1510	0.5430	0.7000	0.4510	0.4900	3.7460	1.1560	1.0260	3.8110	2.6990	2.5270	4.4580
Phenylalanine	1.5140	0.9740		1.1800	1.3140	0.6980	0.5580	2.3240	3.7710	1.3890	0.5790	4.1380	0.5900	0.8730	1.3290	1.1100	0.4970	1.0420	0.4840	0.1870	1.5170	0.9070	0.4630	3.7460	3.8830	2.8880	1.8860
Roseoside	1.2500	1.0110	0.8110	1.4290	1.3230	1.6870	1.3190	2.0990	4.5470	0.8950	0.6510	2.2960	1.0980	0.8450	0.9780	1.6360	0.8920	1.2900	1.0820	0.4300	2.0840	1.7190	1.3140	3.4010	3.8270	2.8760	3.5720
Serine	0.6320	0.3470	0.1810	2.0140	0.8800	0.1790	0.2680	2.7870	0.4510	0.2380	0.3010	1.2140	0.2620	2.4830	0.8380	1.5090	0.8130	2.0390	1.1880	0.6340	0.4090	0.3420	0.7110	0.3400	0.0830	0.2470	0.1480
Sinalbin B	1.0660	2.3620	0.7080		0.7260	1.8260	0.6030	0.2760	5.6740	0.5950	0.4770	1.6740	0.3810	0.5200	0.7130	1.0260	0.5850	1.0570	0.1290	0.0590	0.8020	0.7760		1.5390	0.6560	0.6720	
Threonine	1.1260	0.5800	0.3940	3.1250	0.7810	0.3200	0.2630	2.1680	0.8260	0.2820	0.5150	1.9430	0.7700	2.0320	0.8090	2.0460	0.8950	3.2030	0.6950	0.7560	1.4900	1.0730	0.4600	0.8550	0.3460	0.3780	0.6420
Tyrosine	0.9850	0.5430	0.2050	2.3660	1.1010	0.5740	0.3590	2.7870	2.8910	0.3790	0.3000	2.0340	0.4190	1.2010	0.5940	1.3240	0.4540	0.7420	0.3920	0.6240	0.4860	0.7520	0.6710	1.6200	0.7290	0.7040	1.0350
Valine	1.4840	0.8060	0.5040	3.3420	0.5280	1.1370	0.8890	2.7870	3.0160	0.8660	0.4660	2.4660	0.4660	1.7720	0.7440	1.8310	0.8900	1.6900	0.8720	0.7850	1.0100	0.5140	0.3570	1.1620	0.9750	0.5780	1.1960
MS_METABOLITE_DATA_END
#METABOLITES
METABOLITES_START
metabolite_name	pubchem_id	inchi_key	kegg_id	other_id	other_id_type	ri	ri_type	moverz_quant	
Asparagine	236		C00152					133.06076	
Aspartate	424		C16433					134.04478	
Coniferin			C00761 					341.12419	
Leucine	6106		D00030					130.08735	
Phenylalanine	6140		D00021  					166.08625	
Roseoside	9930064							385.18679	
Serine	5951		C00065					106.04986	
Sinalbin B	656568		C08426					424.03776	
Threonine	6288		D00041 					120.06551	
Tyrosine	6057		C00082					182.08116	
Valine	6287		D00039					118.08625	
METABOLITES_END
#END