#METABOLOMICS WORKBENCH lim_20241024_042947 DATATRACK_ID:5314 STUDY_ID:ST003535 ANALYSIS_ID:AN005806 PROJECT_ID:PR002175
VERSION             	1
CREATED_ON             	October 24, 2024, 9:24 am
#PROJECT
PR:PROJECT_TITLE                 	Arsenic-induced enhancement of diazotrophic recruitment and nitrogen fixation in
PR:PROJECT_TITLE                 	Pteris vittata rhizosphere
PR:PROJECT_SUMMARY               	The enrichment of diazotrophs and resulting nitrogen accumulation promoted
PR:PROJECT_SUMMARY               	hyperaccumulator growth and facilitated phytoremediation. Nonetheless, the
PR:PROJECT_SUMMARY               	regulatory mechanism of hyperaccumulator biological nitrogen fixation has
PR:PROJECT_SUMMARY               	remained elusive. Here, we report the mechanism by which arsenic regulates
PR:PROJECT_SUMMARY               	biological nitrogen fixation in the arsenic-hyperaccumulator Pteris vittata.
PR:PROJECT_SUMMARY               	Field investigations and greenhouse experiments, based on multi-omics
PR:PROJECT_SUMMARY               	approaches, reveal that elevated arsenic stress induces an enrichment of key
PR:PROJECT_SUMMARY               	diazotrophs, enhances plant nitrogen acquisition, and thus improves plant
PR:PROJECT_SUMMARY               	growth. Metabolomic analysis and microfluidic experiments further demonstrate
PR:PROJECT_SUMMARY               	that the upregulation of specific root metabolites plays a crucial role in
PR:PROJECT_SUMMARY               	recruiting key diazotrophic bacteria. These findings highlight the pivotal role
PR:PROJECT_SUMMARY               	of nitrogen-acquisition mechanisms in the arsenic hyperaccumulation of Pteris
PR:PROJECT_SUMMARY               	vittata, and provide valuable insights into the plant stress resistance.
PR:INSTITUTE                     	Institute of Soil and Water Resources and Environmental Science, College of
PR:INSTITUTE                     	Environmental and Resource Sciences, Zhejiang University
PR:LAST_NAME                     	Lin
PR:FIRST_NAME                    	Jiahui
PR:ADDRESS                       	866 Yuhangtang Road, Hangzhou, Zhejiang, 310058, China
PR:EMAIL                         	linjh.link@zju.edu.cn
PR:PHONE                         	18968220088
#STUDY
ST:STUDY_TITLE                   	Arsenic-induced enhancement of diazotrophic recruitment and nitrogen fixation in
ST:STUDY_TITLE                   	Pteris vittata rhizosphere
ST:STUDY_SUMMARY                 	Heavy metal contamination poses an escalating global challenge to soil
ST:STUDY_SUMMARY                 	ecosystems. Hyperaccumulators play a crucial role in environmental remediation
ST:STUDY_SUMMARY                 	and resource recovery. The enrichment of diazotrophs and resulting nitrogen
ST:STUDY_SUMMARY                 	accumulation promoted hyperaccumulator growth and facilitated phytoremediation.
ST:STUDY_SUMMARY                 	Nonetheless, the regulatory mechanism of hyperaccumulator biological nitrogen
ST:STUDY_SUMMARY                 	fixation has remained elusive. Here, we report the mechanism by which arsenic
ST:STUDY_SUMMARY                 	regulates biological nitrogen fixation in the arsenic-hyperaccumulator Pteris
ST:STUDY_SUMMARY                 	vittata. Field investigations and greenhouse experiments, based on multi-omics
ST:STUDY_SUMMARY                 	approaches, reveal that elevated arsenic stress induces an enrichment of key
ST:STUDY_SUMMARY                 	diazotrophs, enhances plant nitrogen acquisition, and thus improves plant
ST:STUDY_SUMMARY                 	growth. Metabolomic analysis and microfluidic experiments further demonstrate
ST:STUDY_SUMMARY                 	that the upregulation of specific root metabolites plays a crucial role in
ST:STUDY_SUMMARY                 	recruiting key diazotrophic bacteria. These findings highlight the pivotal role
ST:STUDY_SUMMARY                 	of nitrogen-acquisition mechanisms in the arsenic hyperaccumulation of Pteris
ST:STUDY_SUMMARY                 	vittata, and provide valuable insights into the plant stress resistance.
ST:INSTITUTE                     	Institute of Soil and Water Resources and Environmental Science, College of
ST:INSTITUTE                     	Environmental and Resource Sciences, Zhejiang University
ST:LAST_NAME                     	Lin
ST:FIRST_NAME                    	Jiahui
ST:ADDRESS                       	866 Yuhangtang Road, Hangzhou, Zhejiang, 310058, China
ST:EMAIL                         	linjh.link@zju.edu.cn
ST:PHONE                         	18968220088
#SUBJECT
SU:SUBJECT_TYPE                  	Plant
SU:SUBJECT_SPECIES               	Pteris vittata
#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           	3	s1	Sample source:Rhizosphere | Treatment:Slight arsenic	Genotype=wild-type; RAW_FILE_NAME=s1.mzXML
SUBJECT_SAMPLE_FACTORS           	4	s2	Sample source:Rhizosphere | Treatment:Slight arsenic	Genotype=wild-type; RAW_FILE_NAME=s2.mzXML
SUBJECT_SAMPLE_FACTORS           	7	s3	Sample source:Rhizosphere | Treatment:Slight arsenic	Genotype=wild-type; RAW_FILE_NAME=s3.mzXML
SUBJECT_SAMPLE_FACTORS           	9	s4	Sample source:Rhizosphere | Treatment:Slight arsenic	Genotype=wild-type; RAW_FILE_NAME=s4.mzXML
SUBJECT_SAMPLE_FACTORS           	11	s5	Sample source:Rhizosphere | Treatment:Slight arsenic	Genotype=wild-type; RAW_FILE_NAME=s5.mzXML
SUBJECT_SAMPLE_FACTORS           	12	s6	Sample source:Rhizosphere | Treatment:Slight arsenic	Genotype=wild-type; RAW_FILE_NAME=s6.mzXML
SUBJECT_SAMPLE_FACTORS           	13	s7	Sample source:Rhizosphere | Treatment:Slight arsenic	Genotype=wild-type; RAW_FILE_NAME=s7.mzXML
SUBJECT_SAMPLE_FACTORS           	14	s8	Sample source:Rhizosphere | Treatment:Slight arsenic	Genotype=wild-type; RAW_FILE_NAME=s8.mzXML
SUBJECT_SAMPLE_FACTORS           	15	s9	Sample source:Rhizosphere | Treatment:Slight arsenic	Genotype=wild-type; RAW_FILE_NAME=s9.mzXML
SUBJECT_SAMPLE_FACTORS           	16	s10	Sample source:Rhizosphere | Treatment:Slight arsenic	Genotype=wild-type; RAW_FILE_NAME=s10.mzXML
SUBJECT_SAMPLE_FACTORS           	3	s11	Sample source:Bulk soil | Treatment:Slight arsenic	Genotype=wild-type; RAW_FILE_NAME=s11.mzXML
SUBJECT_SAMPLE_FACTORS           	4	s12	Sample source:Bulk soil | Treatment:Slight arsenic	Genotype=wild-type; RAW_FILE_NAME=s12.mzXML
SUBJECT_SAMPLE_FACTORS           	7	s13	Sample source:Bulk soil | Treatment:Slight arsenic	Genotype=wild-type; RAW_FILE_NAME=s13.mzXML
SUBJECT_SAMPLE_FACTORS           	9	s14	Sample source:Bulk soil | Treatment:Slight arsenic	Genotype=wild-type; RAW_FILE_NAME=s14.mzXML
SUBJECT_SAMPLE_FACTORS           	11	s15	Sample source:Bulk soil | Treatment:Slight arsenic	Genotype=wild-type; RAW_FILE_NAME=s15.mzXML
SUBJECT_SAMPLE_FACTORS           	12	s16	Sample source:Bulk soil | Treatment:Slight arsenic	Genotype=wild-type; RAW_FILE_NAME=s16.mzXML
SUBJECT_SAMPLE_FACTORS           	13	s17	Sample source:Bulk soil | Treatment:Slight arsenic	Genotype=wild-type; RAW_FILE_NAME=s17.mzXML
SUBJECT_SAMPLE_FACTORS           	14	s18	Sample source:Bulk soil | Treatment:Slight arsenic	Genotype=wild-type; RAW_FILE_NAME=s18.mzXML
SUBJECT_SAMPLE_FACTORS           	15	s19	Sample source:Bulk soil | Treatment:Slight arsenic	Genotype=wild-type; RAW_FILE_NAME=s19.mzXML
SUBJECT_SAMPLE_FACTORS           	16	s20	Sample source:Bulk soil | Treatment:Slight arsenic	Genotype=wild-type; RAW_FILE_NAME=s20.mzXML
SUBJECT_SAMPLE_FACTORS           	G1	s21	Sample source:Rhizosphere | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=s21.mzXML
SUBJECT_SAMPLE_FACTORS           	G2	s22	Sample source:Rhizosphere | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=s22.mzXML
SUBJECT_SAMPLE_FACTORS           	G3	s23	Sample source:Rhizosphere | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=s23.mzXML
SUBJECT_SAMPLE_FACTORS           	G4	s24	Sample source:Rhizosphere | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=s24.mzXML
SUBJECT_SAMPLE_FACTORS           	G5	s25	Sample source:Rhizosphere | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=s25.mzXML
SUBJECT_SAMPLE_FACTORS           	G6	s26	Sample source:Rhizosphere | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=s26.mzXML
SUBJECT_SAMPLE_FACTORS           	G7	s27	Sample source:Rhizosphere | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=s27.mzXML
SUBJECT_SAMPLE_FACTORS           	G8	s28	Sample source:Rhizosphere | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=s28.mzXML
SUBJECT_SAMPLE_FACTORS           	G9	s29	Sample source:Rhizosphere | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=s29.mzXML
SUBJECT_SAMPLE_FACTORS           	G10	s30	Sample source:Rhizosphere | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=s30.mzXML
SUBJECT_SAMPLE_FACTORS           	G11	s31	Sample source:Rhizosphere | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=s31.mzXML
SUBJECT_SAMPLE_FACTORS           	G12	s32	Sample source:Rhizosphere | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=s32.mzXML
SUBJECT_SAMPLE_FACTORS           	G13	s33	Sample source:Rhizosphere | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=s33.mzXML
SUBJECT_SAMPLE_FACTORS           	G14	s34	Sample source:Rhizosphere | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=s34.mzXML
SUBJECT_SAMPLE_FACTORS           	G15	s35	Sample source:Rhizosphere | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=s35.mzXML
SUBJECT_SAMPLE_FACTORS           	G1	s36	Sample source:Bulk soil | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=s36.mzXML
SUBJECT_SAMPLE_FACTORS           	G2	s37	Sample source:Bulk soil | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=s37.mzXML
SUBJECT_SAMPLE_FACTORS           	G3	s38	Sample source:Bulk soil | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=s38.mzXML
SUBJECT_SAMPLE_FACTORS           	G4	s39	Sample source:Bulk soil | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=s39.mzXML
SUBJECT_SAMPLE_FACTORS           	G5	s40	Sample source:Bulk soil | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=s40.mzXML
SUBJECT_SAMPLE_FACTORS           	G6	s41	Sample source:Bulk soil | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=s41.mzXML
SUBJECT_SAMPLE_FACTORS           	G7	s42	Sample source:Bulk soil | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=s42.mzXML
SUBJECT_SAMPLE_FACTORS           	G8	s43	Sample source:Bulk soil | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=s43.mzXML
SUBJECT_SAMPLE_FACTORS           	G9	s44	Sample source:Bulk soil | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=s44.mzXML
SUBJECT_SAMPLE_FACTORS           	G10	s45	Sample source:Bulk soil | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=s45.mzXML
SUBJECT_SAMPLE_FACTORS           	G11	s46	Sample source:Bulk soil | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=s46.mzXML
SUBJECT_SAMPLE_FACTORS           	G12	s47	Sample source:Bulk soil | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=s47.mzXML
SUBJECT_SAMPLE_FACTORS           	G13	s48	Sample source:Bulk soil | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=s48.mzXML
SUBJECT_SAMPLE_FACTORS           	G14	s49	Sample source:Bulk soil | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=s49.mzXML
SUBJECT_SAMPLE_FACTORS           	G15	s50	Sample source:Bulk soil | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=s50.mzXML
SUBJECT_SAMPLE_FACTORS           	3	root1	Sample source:Root | Treatment:Slight arsenic	Genotype=wild-type; RAW_FILE_NAME=root1.mzXML
SUBJECT_SAMPLE_FACTORS           	4	root2	Sample source:Root | Treatment:Slight arsenic	Genotype=wild-type; RAW_FILE_NAME=root2.mzXML
SUBJECT_SAMPLE_FACTORS           	7	root3	Sample source:Root | Treatment:Slight arsenic	Genotype=wild-type; RAW_FILE_NAME=root3.mzXML
SUBJECT_SAMPLE_FACTORS           	9	root4	Sample source:Root | Treatment:Slight arsenic	Genotype=wild-type; RAW_FILE_NAME=root4.mzXML
SUBJECT_SAMPLE_FACTORS           	11	root5	Sample source:Root | Treatment:Slight arsenic	Genotype=wild-type; RAW_FILE_NAME=root5.mzXML
SUBJECT_SAMPLE_FACTORS           	12	root6	Sample source:Root | Treatment:Slight arsenic	Genotype=wild-type; RAW_FILE_NAME=root6.mzXML
SUBJECT_SAMPLE_FACTORS           	13	root7	Sample source:Root | Treatment:Slight arsenic	Genotype=wild-type; RAW_FILE_NAME=root7.mzXML
SUBJECT_SAMPLE_FACTORS           	14	root8	Sample source:Root | Treatment:Slight arsenic	Genotype=wild-type; RAW_FILE_NAME=root8.mzXML
SUBJECT_SAMPLE_FACTORS           	15	root9	Sample source:Root | Treatment:Slight arsenic	Genotype=wild-type; RAW_FILE_NAME=root9.mzXML
SUBJECT_SAMPLE_FACTORS           	16	root10	Sample source:Root | Treatment:Slight arsenic	Genotype=wild-type; RAW_FILE_NAME=root10.mzXML
SUBJECT_SAMPLE_FACTORS           	G1	rootG1	Sample source:Root | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=rootG1.mzXML
SUBJECT_SAMPLE_FACTORS           	G2	rootG2	Sample source:Root | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=rootG2.mzXML
SUBJECT_SAMPLE_FACTORS           	G3	rootG3	Sample source:Root | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=rootG3.mzXML
SUBJECT_SAMPLE_FACTORS           	G4	rootG4	Sample source:Root | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=rootG4.mzXML
SUBJECT_SAMPLE_FACTORS           	G5	rootG5	Sample source:Root | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=rootG5.mzXML
SUBJECT_SAMPLE_FACTORS           	G6	rootG6	Sample source:Root | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=rootG6.mzXML
SUBJECT_SAMPLE_FACTORS           	G7	rootG7	Sample source:Root | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=rootG7.mzXML
SUBJECT_SAMPLE_FACTORS           	G8	rootG8	Sample source:Root | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=rootG8.mzXML
SUBJECT_SAMPLE_FACTORS           	G9	rootG9	Sample source:Root | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=rootG9.mzXML
SUBJECT_SAMPLE_FACTORS           	G10	rootG10	Sample source:Root | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=rootG10.mzXML
SUBJECT_SAMPLE_FACTORS           	G11	rootG11	Sample source:Root | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=rootG11.mzXML
SUBJECT_SAMPLE_FACTORS           	G12	rootG12	Sample source:Root | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=rootG12.mzXML
SUBJECT_SAMPLE_FACTORS           	G13	rootG13	Sample source:Root | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=rootG13.mzXML
SUBJECT_SAMPLE_FACTORS           	G14	rootG14	Sample source:Root | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=rootG14.mzXML
SUBJECT_SAMPLE_FACTORS           	G15	rootG15	Sample source:Root | Treatment:Severe arsenic	Genotype=wild-type; RAW_FILE_NAME=rootG15.mzXML
#COLLECTION
CO:COLLECTION_SUMMARY            	To collect rhizosphere metabolites, roots were gently shaken to remove loosely
CO:COLLECTION_SUMMARY            	adhering soil, followed by the careful collection of the closely adhering soil.
CO:COLLECTION_SUMMARY            	The collected rhizosphere soil was then freeze-dried and pulverized in a grinder
CO:COLLECTION_SUMMARY            	with glass beads (55 Hz, 2 min). Subsequently, 5 g of rhizospheric soil was
CO:COLLECTION_SUMMARY            	transferred into a sterile centrifuge tube containing 25 mL of a pre-cooled (4
CO:COLLECTION_SUMMARY            	℃) mixed solution of acetonitrile: methanol: H2O (2:2:1, v/v/v). The mixture
CO:COLLECTION_SUMMARY            	underwent two rounds of vortexing for 30 s each on a BE-96 Vortex mixer, coupled
CO:COLLECTION_SUMMARY            	with 30-min ultrasonication at 60 Hz on a KW-100TDV ultrasonic cleaner.
CO:COLLECTION_SUMMARY            	Following centrifugation at 10,000 × g for 10 min at 4 ℃ using a H1850-R
CO:COLLECTION_SUMMARY            	refrigerated centrifuge, the supernatant was filtered through a 0.2-μm sterile
CO:COLLECTION_SUMMARY            	membrane filter (Millipore, Bedford, USA) to isolate the rhizosphere
CO:COLLECTION_SUMMARY            	metabolites. The metabolites were then concentrated into a 2-mL centrifuge tube,
CO:COLLECTION_SUMMARY            	dried into a powder, and stored at -80 ℃. This procedure was also applied for
CO:COLLECTION_SUMMARY            	the collection of bulk soil metabolites.
CO:SAMPLE_TYPE                   	Root, Rhizosphere, Soil
#TREATMENT
TR:TREATMENT_SUMMARY             	wild Pteris vittata plants with intact roots and rhizosphere soils and adjacent
TR:TREATMENT_SUMMARY             	bulk soils (0.3 meters from the root) were also collected from the field sites
TR:TREATMENT_SUMMARY             	with low-As (<1000 ppm, n=10) and high-As stress (>1000 ppm, n=15),
TR:TREATMENT_SUMMARY             	respectively, in Wenshan, Yunnan (22°54'02.6"N, 104°23'08.4"E). The in-situ
TR:TREATMENT_SUMMARY             	concentrations of heavy metals (metalloids) were evaluated using the Vanta™
TR:TREATMENT_SUMMARY             	Handheld XRF Series (Tokyo, Japan). In the laboratory, the rhizospheric soils
TR:TREATMENT_SUMMARY             	and roots from Pteris vittata samples, alongside the collected bulk soils, were
TR:TREATMENT_SUMMARY             	processed for subsequent metagenomic and metabolomic analyses.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Equal amounts of preprocessed metabolome powder were re-dissolved by the precise
SP:SAMPLEPREP_SUMMARY            	addition of 300 μL of an acetonitrile solution containing 4 ppm
SP:SAMPLEPREP_SUMMARY            	2-amino-3-(2-chlorophenyl)-propionic acid, prepared with 0.1% formic acid (1:9,
SP:SAMPLEPREP_SUMMARY            	v/v; stored at 4 ℃). The supernatant was then filtered through 0.2-μm
SP:SAMPLEPREP_SUMMARY            	membranes (Millipore, Bedford, USA) and transferred into vials for further
SP:SAMPLEPREP_SUMMARY            	detection.
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Thermo Vanquish
CH:COLUMN_NAME                   	Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um)
CH:SOLVENT_A                     	100% acetonitrile
CH:SOLVENT_B                     	100% water; 5 mM ammonium formate
CH:FLOW_GRADIENT                 	2:98 (A/B, v/v) at 0 min, 2:98 (A/B, v/v) at 1.0 min, 50:50 (A/B, v/v) at 9.0
CH:FLOW_GRADIENT                 	min, 98:2 (A/B, v/v) at 12.0 min, 98:2 (A/B, v/v) at 13.5 min, 2:98 (A/B, v/v)
CH:FLOW_GRADIENT                 	at 14.0 min, 2:98 (A/B, v/v) at 17.0 min
CH:FLOW_RATE                     	0.25 mL/min
CH:COLUMN_TEMPERATURE            	40
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Thermo Q Exactive HF-X Orbitrap
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	POSITIVE
MS:MS_COMMENTS                   	In untargeted metabolomics analysis, the raw MS data were converted to mzXML
MS:MS_COMMENTS                   	format by MSConvert in ProteoWizard software package (v3.0) and processed by
MS:MS_COMMENTS                   	XCMS70 (v3.2) for peak picking and alignment. The XCMS settings were as follows:
MS:MS_COMMENTS                   	method = “centWave”, ppm = 15, peakwidth = c(5,30), mzwid = 0.015, mzdiff =
MS:MS_COMMENTS                   	0.01, and bw = 2.
MS:MS_RESULTS_FILE               	ST003535_AN005806_Results.txt	UNITS:Peak area	Has m/z:Yes	Has RT:Yes	RT units:Seconds
#END