{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST000353","ANALYSIS_ID":"AN000575","VERSION":"1","CREATED_ON":"February 23, 2016, 9:42 pm"},

"PROJECT":{"PROJECT_TITLE":"The Development of Metabolomic Markers in Bermudagrass (Cynodon spp.) for Sting Nematode (Belonolaimus longicaudatus) Response","PROJECT_TYPE":"The majority of golf courses in Florida experience damage from plant-parasitic nematodes and more than 80% of golf courses in Florida are at risk for nematode-related damage (Crow, 2005). With limited availability of nematicides and lack of tolerant cultivars, turf managers need new genetically resistant cultivars. Use of resistant or tolerant grasses is the most efficient, environmentally friendly and, in the long run, least costly component of IPM.","PROJECT_SUMMARY":"Current screening of new genotypes for nematode resistance is labor- and time-intensive, with three months required soley for nematode development on test plants. We intend to develop metabolomic markers or a signature pattern associated with nematode resistance or tolerance. Screening plants for these markers or a signature pattern of would greatly speed up the screening process and development of resistant cultivars.","INSTITUTE":"University of Florida","DEPARTMENT":"Agronomy","LABORATORY":"Turfgrass Breeding","LAST_NAME":"Kenworthy","FIRST_NAME":"Kevin","ADDRESS":"2005 SW 23rd Street, Gainesville FL 32611","EMAIL":"nbenda@ufl.edu","PHONE":"352-792-4561"},

"STUDY":{"STUDY_TITLE":"The Development of Metabolomic Markers in African Bermudagrass (C. transvaalensis) for Sting Nematode (Belonolaimus longicaudatus) Response","STUDY_TYPE":"Disease response in terms of nematode reproduction and root weight","STUDY_SUMMARY":"The objective of the proposed pilot study is to identify metabolites up- and down-regulated in African bermudagrass that are tolerant and sensitive to the sting nematode and develop metabolomic markers for the highest expressed metabolites associated with tolerance. Future work will include additional accessions and species of bermudagrass, and testing under field conditions. Bermudagrass accessions identified as tolerant or sensitive by Pang et al. (2011) will be assessed under controlled greenhouse conditions to identify metabolites linked to sting nematode tolerance. Nematode response will be quantified through determination of root length and weight and the number of nematodes present 136 days after inoculation. Higher root length and weight indicate tolerance or resistance. Higher nematode counts indicate greater reproduction (i.e. a susceptible plant), while lower counts indicate that the accession may have some resistance. Metabolites from root tissue of these accessions will be compared to identify those associated with tolerance/resistance, and those that are associated with nematode infestation by comparing inoculated plants to uninoculated controls. Metabolomic markers will then be developed for the metabolites associated with tolerance/resistance. These markers will be used to guide future screening of bermudagrass accessions for breeding nematode-tolerant or -resistant varieties.","INSTITUTE":"University of Florida","DEPARTMENT":"SECIM","LABORATORY":"Turfgrass Breeding","LAST_NAME":"Benda","FIRST_NAME":"Nicole","ADDRESS":"2005 SW 23rd St","EMAIL":"nbenda@ufl.edu","PHONE":"352-792-4561","NUM_GROUPS":"3","TOTAL_SUBJECTS":"42"},

"SUBJECT":{"SUBJECT_TYPE":"Plant","SUBJECT_SPECIES":"Cynodon transvaalensis","GENOTYPE_STRAIN":"AB03, AB33, and AB39","AGE_OR_AGE_RANGE":"4 month old plant clones","SUBJECT_COMMENTS":"(African Bermudagrass,C. transvaalensis)"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"AB03_I_1",
"Factors":{"Plant genotype":"AB03","Nematode inoculation":"inoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB03_I_2",
"Factors":{"Plant genotype":"AB03","Nematode inoculation":"inoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB03_I_4",
"Factors":{"Plant genotype":"AB03","Nematode inoculation":"inoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB03_I_5",
"Factors":{"Plant genotype":"AB03","Nematode inoculation":"inoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB03_I_7",
"Factors":{"Plant genotype":"AB03","Nematode inoculation":"inoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB03_I_10",
"Factors":{"Plant genotype":"AB03","Nematode inoculation":"inoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB03_I_13",
"Factors":{"Plant genotype":"AB03","Nematode inoculation":"inoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB03_U_3",
"Factors":{"Plant genotype":"AB03","Nematode inoculation":"uninoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB03_U_4",
"Factors":{"Plant genotype":"AB03","Nematode inoculation":"uninoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB03_U_6",
"Factors":{"Plant genotype":"AB03","Nematode inoculation":"uninoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB03_U_7",
"Factors":{"Plant genotype":"AB03","Nematode inoculation":"uninoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB03_U_8",
"Factors":{"Plant genotype":"AB03","Nematode inoculation":"uninoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB03_U_11",
"Factors":{"Plant genotype":"AB03","Nematode inoculation":"uninoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB03_U_14",
"Factors":{"Plant genotype":"AB03","Nematode inoculation":"uninoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB33_I_1",
"Factors":{"Plant genotype":"AB33","Nematode inoculation":"inoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB33_I_2",
"Factors":{"Plant genotype":"AB33","Nematode inoculation":"inoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB33_I_3",
"Factors":{"Plant genotype":"AB33","Nematode inoculation":"inoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB33_I_5",
"Factors":{"Plant genotype":"AB33","Nematode inoculation":"inoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB33_I_8",
"Factors":{"Plant genotype":"AB33","Nematode inoculation":"inoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB33_I_10",
"Factors":{"Plant genotype":"AB33","Nematode inoculation":"inoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB33_I_11",
"Factors":{"Plant genotype":"AB33","Nematode inoculation":"inoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB33_U_2",
"Factors":{"Plant genotype":"AB33","Nematode inoculation":"uninoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB33_U_4",
"Factors":{"Plant genotype":"AB33","Nematode inoculation":"uninoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB33_U_7",
"Factors":{"Plant genotype":"AB33","Nematode inoculation":"uninoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB33_U_8",
"Factors":{"Plant genotype":"AB33","Nematode inoculation":"uninoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB33_U_9",
"Factors":{"Plant genotype":"AB33","Nematode inoculation":"uninoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB33_U_11",
"Factors":{"Plant genotype":"AB33","Nematode inoculation":"uninoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB33_U_13",
"Factors":{"Plant genotype":"AB33","Nematode inoculation":"uninoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB39_I_1",
"Factors":{"Plant genotype":"AB39","Nematode inoculation":"inoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB39_I_3",
"Factors":{"Plant genotype":"AB39","Nematode inoculation":"inoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB39_I_4",
"Factors":{"Plant genotype":"AB39","Nematode inoculation":"inoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB39_I_8",
"Factors":{"Plant genotype":"AB39","Nematode inoculation":"inoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB39_I_9",
"Factors":{"Plant genotype":"AB39","Nematode inoculation":"inoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB39_I_11",
"Factors":{"Plant genotype":"AB39","Nematode inoculation":"inoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB39_I_12",
"Factors":{"Plant genotype":"AB39","Nematode inoculation":"inoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB39_U_2",
"Factors":{"Plant genotype":"AB39","Nematode inoculation":"uninoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB39_U_3",
"Factors":{"Plant genotype":"AB39","Nematode inoculation":"uninoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB39_U_5",
"Factors":{"Plant genotype":"AB39","Nematode inoculation":"uninoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB39_U_6",
"Factors":{"Plant genotype":"AB39","Nematode inoculation":"uninoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB39_U_7",
"Factors":{"Plant genotype":"AB39","Nematode inoculation":"uninoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB39_U_9",
"Factors":{"Plant genotype":"AB39","Nematode inoculation":"uninoc"}
},
{
"Subject ID":"-",
"Sample ID":"AB39_U_10",
"Factors":{"Plant genotype":"AB39","Nematode inoculation":"uninoc"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Rinsed roots were cut off the plant, placed into paper envelopes, and immediately placed into liquid nitrogen. Roots were stored at -80°C until freeze-drying. Freeze-dried roots were held at room temperature in ziploc bags with silica gel packets. They were then frozen in liquid nitrogen and ground to a powder in a tissue homogenizer with ball bearings (Geno/Grinder 2010), and then stored in 2 ml snap-cap plastic tubes at room temperature.","SAMPLE_TYPE":"plant root tissue","COLLECTION_METHOD":"whole plant destruction","COLLECTION_LOCATION":"greenhouse","COLLECTION_FREQUENCY":"once","COLLECTION_DURATION":"1-2 minutes to cut off and rinse the roots prior to placement into liquid nitrogen","COLLECTION_TIME":"136 days after inoculation","VOLUMEORAMOUNT_COLLECTED":"0.2-0.5 grams of root tissue, dry weight","STORAGE_CONDITIONS":"1)-80C until freeze-drying. 2) Held at room temperature in ziploc bags with silica packets. 3) Frozen in liquid nitrogen and ground using a Geno/Grinder 2010. 4) Transferred to 2 mL tubes and held at room temperature.","COLLECTION_VIALS":"Paper coin envelopes","STORAGE_VIALS":"2 ml snap-cap plastic tubes","COLLECTION_TUBE_TEMP":"liquid nitrogen (-196°C)","ADDITIVES":"none"},

"TREATMENT":{"TREATMENT_SUMMARY":"Plants clones propagated by stolons were allowed to establish for 30 days in 4 cm diameter conetainers filled with sand. Fifty sting nematodes (mixed age and gender) were added to each conetainer and allowed to grow and reproduce for 90 days. Nematodes were then rinsed off of roots for quantification, and plant roots were frozen in liquid nitrogen and stored at -80C until they were freeze-dried.Freeze-dried roots were ground to a powder and then submitted for global metabolomic analysis.","TREATMENT":"Biotic","TREATMENT_COMPOUND":"Sting nematode, Belonolaimus longicaudatus","PLANT_GROWTH_SUPPORT":"sand in conetainers- 4 cm wide, 30 cm deep","PLANT_GROWTH_LOCATION":"temperature controlled greenhouse","PLANT_PLOT_DESIGN":"complete randomized design","PLANT_LIGHT_PERIOD":"Natural lighting, ca. 11:13 Light:Dark","PLANT_HUMIDITY":"ca.50%RH","PLANT_TEMP":"25°C","PLANT_WATERING_REGIME":"Twice daily for 5 min","PLANT_NUTRITIONAL_REGIME":"Weekly liquid fertilizer of 24-8-16 (N-P-K)","PLANT_ESTAB_DATE":"09-24-2015","PLANT_HARVEST_DATE":"04-02-2016","PLANT_GROWTH_STAGE":"non-reproductive","PLANT_METAB_QUENCH_METHOD":"liquid nitrogen, followed by storage at -80°C","PLANT_HARVEST_METHOD":"rinsed roots were cut off the plant and placed into paper envelopes","PLANT_STORAGE":"1)-80C until freeze-drying. 2) Held at room temperature in ziploc bags with silica packets. 3) Frozen in liquid nitrogen and ground using a Geno/Grinder 2010. 4) Transferred to 2 mL tubes and held at room temperature."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"none","SAMPLEPREP_PROTOCOL_FILENAME":"Maize_Tissue_sample_prep.pdf"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"See document","CHROMATOGRAPHY_TYPE":"Reversed phase","INSTRUMENT_NAME":"Thermo Scientific-Dionex Ultimate 3000","COLUMN_NAME":"ACE Excel 2 C18-PFP (100 x 2.1mm, 2um)","METHODS_FILENAME":"Metabolomics_LCMSProtocol.pdf;Appendix A - Internal Standard Prep GLCMS.pdf","INTERNAL_STANDARD":"Appendix A - Internal Standard Prep GLCMS.pdf"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS","ANALYSIS_PROTOCOL_FILE":"Metabolomics_LCMSProtocol.pdf"},

"MS":{"INSTRUMENT_NAME":"Thermo Q Exactive Orbitrap","INSTRUMENT_TYPE":"Orbitrap","MS_TYPE":"ESI","ION_MODE":"POSITIVE","MS_COMMENTS":"HESI"},

"MS_METABOLITE_DATA":{
"Units":"peak area",

"Data":[{"Metabolite":"(R)-MALATE","AB33_I_11":"39900000.0000","AB33_I_1":"32500000.0000","AB33_I_10":"69200000.0000","AB33_I_3":"44100000.0000","AB33_I_5":"48900000.0000","AB33_I_2":"44800000.0000","AB03_U_14":"27500000.0000","AB33_I_8":"37500000.0000","AB03_U_6":"28900000.0000","AB03_U_11":"21200000.0000","AB03_U_4":"43500000.0000","AB03_U_7":"26100000.0000","AB03_U_3":"38800000.0000","AB03_I_2":"36700000.0000","AB03_U_8":"38000000.0000","AB03_I_1":"58900000.0000","AB33_U_8":"35000000.0000","AB33_U_2":"53300000.0000","AB03_I_13":"50100000.0000","AB03_I_5":"45400000.0000","AB03_I_7":"37900000.0000","AB33_U_7":"38500000.0000","AB33_U_13":"34700000.0000","AB39_I_3":"33400000.0000","AB03_I_10":"46900000.0000","AB39_I_4":"22500000.0000","AB33_U_9":"40500000.0000","AB33_U_4":"33500000.0000","AB03_I_4":"36200000.0000","AB39_I_1":"39100000.0000","AB39_I_8":"37500000.0000","AB33_U_11":"45600000.0000","AB39_U_9":"22500000.0000","AB39_U_10":"21800000.0000","AB39_I_12":"26200000.0000","AB39_I_11":"28900000.0000","AB39_U_2":"26800000.0000","AB39_U_6":"27800000.0000","AB39_U_3":"29900000.0000","AB39_U_5":"30300000.0000","AB39_U_7":"34300000.0000","AB39_I_9":"49600000.0000"},{"Metabolite":"1-AMINOCYCLOPROPANE-1-CARBOXYLATE","AB33_I_11":"13400000.0000","AB33_I_1":"10400000.0000","AB33_I_10":"10500000.0000","AB33_I_3":"10300000.0000","AB33_I_5":"12800000.0000","AB33_I_2":"12100000.0000","AB03_U_14":"8300000.0000","AB33_I_8":"12500000.0000","AB03_U_6":"5980000.0000","AB03_U_11":"5760000.0000","AB03_U_4":"6720000.0000","AB03_U_7":"7150000.0000","AB03_U_3":"6380000.0000","AB03_I_2":"7170000.0000","AB03_U_8":"7100000.0000","AB03_I_1":"5110000.0000","AB33_U_8":"9880000.0000","AB33_U_2":"11600000.0000","AB03_I_13":"6990000.0000","AB03_I_5":"7490000.0000","AB03_I_7":"7220000.0000","AB33_U_7":"14400000.0000","AB33_U_13":"8340000.0000","AB39_I_3":"8590000.0000","AB03_I_10":"7880000.0000","AB39_I_4":"7130000.0000","AB33_U_9":"9310000.0000","AB33_U_4":"9880000.0000","AB03_I_4":"6990000.0000","AB39_I_1":"8680000.0000","AB39_I_8":"10800000.0000","AB33_U_11":"9760000.0000","AB39_U_9":"8060000.0000","AB39_U_10":"7760000.0000","AB39_I_12":"8130000.0000","AB39_I_11":"9330000.0000","AB39_U_2":"8190000.0000","AB39_U_6":"10000000.0000","AB39_U_3":"11000000.0000","AB39_U_5":"11300000.0000","AB39_U_7":"12400000.0000","AB39_I_9":"12700000.0000"},{"Metabolite":"4-AMINOBUTANOATE","AB33_I_11":"61800000.0000","AB33_I_1":"45800000.0000","AB33_I_10":"80400000.0000","AB33_I_3":"62700000.0000","AB33_I_5":"51800000.0000","AB33_I_2":"46600000.0000","AB03_U_14":"37000000.0000","AB33_I_8":"112000000.0000","AB03_U_6":"23600000.0000","AB03_U_11":"16600000.0000","AB03_U_4":"23700000.0000","AB03_U_7":"25900000.0000","AB03_U_3":"21900000.0000","AB03_I_2":"16400000.0000","AB03_U_8":"27900000.0000","AB03_I_1":"12800000.0000","AB33_U_8":"46500000.0000","AB33_U_2":"55500000.0000","AB03_I_13":"23000000.0000","AB03_I_5":"19400000.0000","AB03_I_7":"13200000.0000","AB33_U_7":"58300000.0000","AB33_U_13":"44500000.0000","AB39_I_3":"40500000.0000","AB03_I_10":"34500000.0000","AB39_I_4":"18200000.0000","AB33_U_9":"36200000.0000","AB33_U_4":"45400000.0000","AB03_I_4":"24400000.0000","AB39_I_1":"9590000.0000","AB39_I_8":"54900000.0000","AB33_U_11":"53700000.0000","AB39_U_9":"23500000.0000","AB39_U_10":"31300000.0000","AB39_I_12":"34900000.0000","AB39_I_11":"33500000.0000","AB39_U_2":"32500000.0000","AB39_U_6":"21900000.0000","AB39_U_3":"39400000.0000","AB39_U_5":"33400000.0000","AB39_U_7":"22300000.0000","AB39_I_9":"46900000.0000"},{"Metabolite":"4-GUANIDINOBUTANOATE","AB33_I_11":"86500000.0000","AB33_I_1":"121000000.0000","AB33_I_10":"116000000.0000","AB33_I_3":"118000000.0000","AB33_I_5":"149000000.0000","AB33_I_2":"82800000.0000","AB03_U_14":"105000000.0000","AB33_I_8":"348000000.0000","AB03_U_6":"43000000.0000","AB03_U_11":"61000000.0000","AB03_U_4":"85600000.0000","AB03_U_7":"39100000.0000","AB03_U_3":"132000000.0000","AB03_I_2":"120000000.0000","AB03_U_8":"59600000.0000","AB03_I_1":"89900000.0000","AB33_U_8":"58300000.0000","AB33_U_2":"72400000.0000","AB03_I_13":"74800000.0000","AB03_I_5":"121000000.0000","AB03_I_7":"109000000.0000","AB33_U_7":"45600000.0000","AB33_U_13":"94800000.0000","AB39_I_3":"30600000.0000","AB03_I_10":"77100000.0000","AB39_I_4":"28000000.0000","AB33_U_9":"89500000.0000","AB33_U_4":"89200000.0000","AB03_I_4":"79100000.0000","AB39_I_1":"39400000.0000","AB39_I_8":"61400000.0000","AB33_U_11":"61000000.0000","AB39_U_9":"76900000.0000","AB39_U_10":"77800000.0000","AB39_I_12":"58500000.0000","AB39_I_11":"124000000.0000","AB39_U_2":"84100000.0000","AB39_U_6":"59400000.0000","AB39_U_3":"78400000.0000","AB39_U_5":"108000000.0000","AB39_U_7":"67400000.0000","AB39_I_9":"74700000.0000"},{"Metabolite":"4-HYDROXY-L-PHENYLGLYCINE","AB33_I_11":"15900000.0000","AB33_I_1":"15700000.0000","AB33_I_10":"16800000.0000","AB33_I_3":"16800000.0000","AB33_I_5":"16100000.0000","AB33_I_2":"17200000.0000","AB03_U_14":"17900000.0000","AB33_I_8":"16800000.0000","AB03_U_6":"129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}

}