{
"METABOLOMICS WORKBENCH":{"STUDY_ID":"ST001927","ANALYSIS_ID":"AN003135","VERSION":"1","CREATED_ON":"02-08-2024"},

"PROJECT":{"PROJECT_TITLE":"Fungal consortium of two Beauveria bassiana strains increases their virulence, growth, and resistance to stress: a metabolomic approach.","PROJECT_TYPE":"Untargeted Metabolomics","PROJECT_SUMMARY":"Entomopathogenic fungi have been successfully used to control agricultural pests. They infect insects by coming into direct contact with their cuticle or when feeding on contaminated leaves or fruits. After contact with the insect, the entomopathogenic fungus penetrates its body cavity, where it grows and colonizes it from within, causing its death The use of two or more microorganisms in a microbial consortium has been increasingly applied in the biological control of diseases and pests. Beauveria bassiana is one of the most widely studied fungal species in biological control, yet little is known about its role in fungal consortiums. In a previous study, our group found that a consortium formed by two strains of B. bassiana had significantly greater biocontrol potential against the polyphagous caterpillars Duponchelia fovealis (Lepidoptera: Crambidae) than either strain on its own. Despite recent developments and growing efforts to better understand fungal metabolism and metabolites, much remains unknown. Metabolomics therefore represents an important field for evaluating the metabolites produced or modified by an organism or its relationship with the environment. In the present study, we aim to use untargeted metabolomics with gas and liquid chromatography coupled to mass spectrometers (GC-MS and LC-MS/MS) to evaluate the metabolic alterations caused by the co-cultivation of these strains and to correlate the metabolites produced by this consortium with the increased mortality in D. fovealis observed previously.","INSTITUTE":"Universidade Federal do Paraná","DEPARTMENT":"Patologia Básica","LABORATORY":"Laboratório de Microbiologia e Biologia Molecular","LAST_NAME":"Stuart","FIRST_NAME":"Andressa","ADDRESS":"Av. Cel. Francisco Heráclito dos Santos, 100, 81530000, Jardim das Américas, Curitiba, Paraná, Brasil","EMAIL":"andressa.katiski@gmail.com","PHONE":"5541991922779","PROJECT_COMMENTS":"Two genetically distinct strains of B. bassiana (Bov 3 and Bov 2) were cultivated in Agar Sabouraud culture medium, both separately and co-cultivated to form a fungal consortium. The metabolomic analysis were performed at the Laboratório de Genética de Plantas Max Feffer facility of the Escola Superior de Agricultura Luiz de Queiroz of the Universidade de São Paulo (ESALQ/USP). Three reads of every biological replicate (five per treatment) were performed, generating fifteen readings for each treatment. Pools of metabolites from each group were created as a quality control.","DOI":"http://dx.doi.org/10.21228/M8D693","CONTRIBUTORS":"Jason Lee Furuie, Thais Regiani Cataldi, Rodrigo Makowiecky Stuart, Maria Aparecida Cassilha Zawadneak, Carlos Alberto Labate, Ida Chapaval Pimentel"},

"STUDY":{"STUDY_TITLE":"Fungal consortium of two Beauveria bassiana strains increases their virulence, growth, and resistance to stress: a metabolomic approach.","STUDY_TYPE":"Untargeted Metabolomics","STUDY_SUMMARY":"Entomopathogenic fungi have been successfully used to control agricultural pests. They infect insects by coming into direct contact with their cuticle or when feeding on contaminated leaves or fruits. After contact with the insect, the entomopathogenic fungus penetrates its body cavity, where it grows and colonizes it from within, causing its death The use of two or more microorganisms in a microbial consortium has been increasingly applied in the biological control of diseases and pests. Beauveria bassiana is one of the most widely studied fungal species in biological control, yet little is known about its role in fungal consortiums. In a previous study, our group found that a consortium formed by two strains of B. bassiana had significantly greater biocontrol potential against the polyphagous caterpillars Duponchelia fovealis (Lepidoptera: Crambidae) than either strain on its own. Despite recent developments and growing efforts to better understand fungal metabolism and metabolites, much remains unknown. Metabolomics therefore represents an important field for evaluating the metabolites produced or modified by an organism or its relationship with the environment. In the present study, we aim to use untargeted metabolomics with gas and liquid chromatography coupled to mass spectrometers (GC-MS and LC-MS/MS) to evaluate the metabolic alterations caused by the co-cultivation of these strains and to correlate the metabolites produced by this consortium with the increased mortality in D. fovealis observed previosly.","INSTITUTE":"Universidade Federal do Paraná","DEPARTMENT":"Patologia Básica","LABORATORY":"Laboratório de Microbiologia e Biologia Molecular","LAST_NAME":"Stuart","FIRST_NAME":"Andressa","ADDRESS":"Av. Cel. Francisco Heráclito dos Santos, 100, 81530000, Jardim das Américas, Curitiba, Paraná, Brasil","EMAIL":"andressa.katiski@gmail.com","PHONE":"5541991922779","SUBMIT_DATE":"2021-09-29"},

"SUBJECT":{"SUBJECT_TYPE":"Fungi","SUBJECT_SPECIES":"Beauveria bassiana","GENOTYPE_STRAIN":"GenBank: KU751847; KU751848"},
"SUBJECT_SAMPLE_FACTORS":[
{
"Subject ID":"-",
"Sample ID":"Bov2_1",
"Factors":{"Fungal species":"Beauveria bassiana Strain Bov2"},
"Additional sample data":{"RAW_FILE_NAME":"Bov2_1","RAW_FILE_NAME":"Bov2_1_R001 Bov2_1_R002 Bov2_1_R003","RAW_FILE_NAME":"Bov2_1_R001_NEG Bov2_1_R002_NEG Bov2_1_R003_NEG","Group":"Bov2"}
},
{
"Subject ID":"-",
"Sample ID":"Bov2_2",
"Factors":{"Fungal species":"Beauveria bassiana Strain Bov2"},
"Additional sample data":{"RAW_FILE_NAME":"Bov2_2","RAW_FILE_NAME":"Bov2_2_R001 Bov2_2_R002 Bov2_2_R003","RAW_FILE_NAME":"Bov2_2_R001_NEG Bov2_2_R002_NEG Bov2_2_R003_NEG","Group":"Bov2"}
},
{
"Subject ID":"-",
"Sample ID":"Bov2_3",
"Factors":{"Fungal species":"Beauveria bassiana Strain Bov2"},
"Additional sample data":{"RAW_FILE_NAME":"Bov2_3","RAW_FILE_NAME":"Bov2_3_R001 Bov2_3_R002 Bov2_3_R003","RAW_FILE_NAME":"Bov2_3_R001_NEG Bov2_3_R003_NEG Bov2_3_R003_NEG","Group":"Bov2"}
},
{
"Subject ID":"-",
"Sample ID":"Bov2_4",
"Factors":{"Fungal species":"Beauveria bassiana Strain Bov2"},
"Additional sample data":{"RAW_FILE_NAME":"Bov2_4","RAW_FILE_NAME":"Bov2_4_R001 Bov2_4_R002 Bov2_4_R003","RAW_FILE_NAME":"Bov2_4_R001_NEG Bov2_4_R002_NEG Bov2_4_R003_NEG","Group":"Bov2"}
},
{
"Subject ID":"-",
"Sample ID":"Bov2_5",
"Factors":{"Fungal species":"Beauveria bassiana Strain Bov2"},
"Additional sample data":{"RAW_FILE_NAME":"Bov2_5","RAW_FILE_NAME":"Bov2_5_R001 Bov2_5_R002 Bov2_5_R003","RAW_FILE_NAME":"Bov2_5_R001_NEG Bov2_5_R002_NEG Bov2_5_R003_NEG","Group":"Bov2"}
},
{
"Subject ID":"-",
"Sample ID":"Bov3_1",
"Factors":{"Fungal species":"Beauveria bassiana Strain Bov3"},
"Additional sample data":{"RAW_FILE_NAME":"Bov3_1","RAW_FILE_NAME":"Bov3_1_R001 Bov3_1_R002 Bov3_1_R003","RAW_FILE_NAME":"Bov3_1_R001_NEG Bov3_1_R002_NEG Bov3_1_R003_NEG","Group":"Bov3"}
},
{
"Subject ID":"-",
"Sample ID":"Bov3_2",
"Factors":{"Fungal species":"Beauveria bassiana Strain Bov3"},
"Additional sample data":{"RAW_FILE_NAME":"Bov3_2","RAW_FILE_NAME":"Bov3_2_R001 Bov3_2_R002 Bov3_2_R003","RAW_FILE_NAME":"Bov3_2_R001_NEG Bov3_2_R002_NEG Bov3_2_R003_NEG","Group":"Bov3"}
},
{
"Subject ID":"-",
"Sample ID":"Bov3_3",
"Factors":{"Fungal species":"Beauveria bassiana Strain Bov3"},
"Additional sample data":{"RAW_FILE_NAME":"Bov3_3","RAW_FILE_NAME":"Bov3_3_R001 Bov3_3_R002 Bov3_3_R003","RAW_FILE_NAME":"Bov3_3_R001_NEG Bov3_3_R002_NEG Bov3_3_R003_NEG","Group":"Bov3"}
},
{
"Subject ID":"-",
"Sample ID":"Bov3_4",
"Factors":{"Fungal species":"Beauveria bassiana Strain Bov3"},
"Additional sample data":{"RAW_FILE_NAME":"Bov3_4","RAW_FILE_NAME":"Bov3_4_R001 Bov3_4_R002 Bov3_4_R003","RAW_FILE_NAME":"Bov3_4_R001_NEG Bov3_4_R002_NEG Bov3_4_R003_NEG","Group":"Bov3"}
},
{
"Subject ID":"-",
"Sample ID":"Bov3_5",
"Factors":{"Fungal species":"Beauveria bassiana Strain Bov3"},
"Additional sample data":{"RAW_FILE_NAME":"Bov3_5","RAW_FILE_NAME":"Bov3_5_R001 Bov3_5_R002 Bov3_5_R003","RAW_FILE_NAME":"Bov3_5_R001_NEG Bov3_5_R002_NEG Bov3_5_R003_NEG","Group":"Bov3"}
},
{
"Subject ID":"-",
"Sample ID":"Consortium_1",
"Factors":{"Fungal species":"Consortium formed by Bov2 and Bov3 strains"},
"Additional sample data":{"RAW_FILE_NAME":"Cons_1","RAW_FILE_NAME":"Cons_1_R001 Cons_1_R002 Cons_1_R003","RAW_FILE_NAME":"Cons_1_R001_NEG Cons_1_R002_NEG Cons_1_R003_NEG","Group":"Cons"}
},
{
"Subject ID":"-",
"Sample ID":"Consortium_2",
"Factors":{"Fungal species":"Consortium formed by Bov2 and Bov3 strains"},
"Additional sample data":{"RAW_FILE_NAME":"Cons_2","RAW_FILE_NAME":"Cons_2_R001 Cons_2_R002 Cons_2_R003","RAW_FILE_NAME":"Cons_2_R001_NEG Cons_2_R002_NEG Cons_2_R003_NEG","Group":"Cons"}
},
{
"Subject ID":"-",
"Sample ID":"Consortium_3",
"Factors":{"Fungal species":"Consortium formed by Bov2 and Bov3 strains"},
"Additional sample data":{"RAW_FILE_NAME":"Cons_3","RAW_FILE_NAME":"Cons_3_R001 Cons_3_R002 Cons_3_R003","RAW_FILE_NAME":"Cons_3_R001_NEG Cons_3_R002_NEG Cons_3_R003_NEG","Group":"Cons"}
},
{
"Subject ID":"-",
"Sample ID":"Consortium_4",
"Factors":{"Fungal species":"Consortium formed by Bov2 and Bov3 strains"},
"Additional sample data":{"RAW_FILE_NAME":"Cons_4","RAW_FILE_NAME":"Cons_4_R001 Cons_4_R002 Cons_4_R003","RAW_FILE_NAME":"Cons_4_R001_NEG Cons_4_R002_NEG Cons_4_R003_NEG","Group":"Cons"}
},
{
"Subject ID":"-",
"Sample ID":"Consortium_5",
"Factors":{"Fungal species":"Consortium formed by Bov2 and Bov3 strains"},
"Additional sample data":{"RAW_FILE_NAME":"Cons_5","RAW_FILE_NAME":"Cons_5_R001 Cons_5_R002 Cons_5_R003","RAW_FILE_NAME":"Cons_5_R001_NEG Cons_5_R002_NEG Cons_5_R003_NEG","Group":"Cons"}
}
],
"COLLECTION":{"COLLECTION_SUMMARY":"Two genetically distinct strains of Beauveria bassiana (Bov 3 and Bov 2) were cultivated both separately and co-cultivated to form a fungal consortium. After the colonies had grown, the mycelium of each treatment (Bov 2, Bov 3, and the fungal consortium) was scraped from the culture medium with a spatula and then macerated separately in liquid nitrogen (N2)","COLLECTION_PROTOCOL_FILENAME":"MetaboliteExtraction","SAMPLE_TYPE":"Fungal mycelium","STORAGE_CONDITIONS":"-80℃"},

"TREATMENT":{"TREATMENT_SUMMARY":"Two genetically distinct strains of B. bassiana (Bov 3 and Bov 2) were cultivated in Petri dishes containing Agar Sabouraud culture medium, both separately and co-cultivated to form a fungal consortium. The cultures were incubated in the dark in a biological oxygen demand (BOD) oven for 14 days at 28°C."},

"SAMPLEPREP":{"SAMPLEPREP_SUMMARY":"Extraction was performed in microtubes, from 200 mg of fungal macerate to which 1 mL of 6:2:2 methanol:chloroform:water ice-cold extraction solution was added. These extraction microtubes were vigorously vortexed and placed in an ultrasonic low-temperature bath at 20 Hz s-1 for 15 min. The samples were then centrifuged (Eppendorf, Germany) at 4°C for 10 min at 14,000 rpm. Then, the supernatant was filtered using a 0.22 μm Whatman® filter (Merck, Germany) and transferred to a chromatographic vial where the extracts were lyophilized (Thermo Fischer Scientific, MA, USA) until completely dry. Finally, the lyophilized samples were resuspended in 200 μL of extraction solution and aliquoted for use in the GC-MS and LC-MS/MS.","PROCESSING_STORAGE_CONDITIONS":"-80℃","EXTRACT_STORAGE":"-80℃"},

"CHROMATOGRAPHY":{"CHROMATOGRAPHY_SUMMARY":"Negative mode (.raw)","METHODS_FILENAME":"LCMSMS","INSTRUMENT_NAME":"Waters Acquity UPLC","COLUMN_NAME":"Waters Acquity UPLC HSS (100 x 2.1mm, 1.7um)","COLUMN_TEMPERATURE":"35 ºC","FLOW_GRADIENT":"95% solvent A and 5% B. The gradient increased linearly to 75% A and 25% B over the next 6 min. The polarity was reversed to 25% A and 75% B for 6 min, and finally 5% A and 95% B for 1 min","FLOW_RATE":"0.5 mL·min-1","SOLVENT_A":"Water; formic acid","SOLVENT_B":"100% acetonitrile; formic acid.","CHROMATOGRAPHY_TYPE":"Reversed phase"},

"ANALYSIS":{"ANALYSIS_TYPE":"MS"},

"MS":{"INSTRUMENT_NAME":"Waters Ultima QTOF","INSTRUMENT_TYPE":"QTOF","MS_TYPE":"ESI","MS_COMMENTS":"Generated data were pre-processed using MassLynx 4.1 software","ION_MODE":"NEGATIVE","CAPILLARY_VOLTAGE":"3 kV","SOURCE_TEMPERATURE":"150 ºC","DESOLVATION_GAS_FLOW":"550 L/hr.","MS_RESULTS_FILE":"ST001927_AN003135_Results.txt UNITS:Relative intensity Has m/z:Yes Has RT:Yes RT units:Minutes"}

}