Summary of Study ST001990
This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001264. The data can be accessed directly via it's Project DOI: 10.21228/M8B39F This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.
| Study ID | ST001990 |
| Study Title | Metabolomics of the interaction between a consortium of entomopathogenic fungi and their target insect: mechanisms of attack and survival |
| Study Type | Untargeted Metabolomics |
| Study Summary | One of the most concerning pests that attack strawberries in Brazil is Duponchelia fovealis, a non-native moth with no registered control methods to date. Our group recently observed that a fungal consortium formed by two strains of Beauveria bassiana increased the mortality of D. fovealis more than inoculation with each strain on its own. However, the molecular interaction between the fungal consortium and the caterpillars is unknown, raising several questions about the enhanced pest control observed. Furthermore, concerns over the emergency of resistance and the selection for resistance to chemical and biological products that are constantly applied in agriculture highlight the need for careful examination of novel pest control methods. Thus, in this work, we sought to pioneer the evaluation of the molecular interaction between a fungal consortium of B. bassiana and D. fovealis caterpillars. We aimed to understand the biocontrol process involved in this interaction and the defense system of the caterpillar. Therefore, seven days after D. fovealis caterpillars were inoculated with the B. bassiana consortium, the dead and surviving caterpillars were analyzed using GC-MS and LC-MS/MS. |
| Institute | Universidade Federal do Paraná |
| Department | Patologia Básica |
| Laboratory | Laboratório de Microbiologia e Biologia Molecular |
| Last Name | Katiski da Costa Stuart |
| First Name | Andressa |
| Address | Av. Cel. Francisco Heráclito dos Santos, 100, Curitiba, Paraná, 81530-000, Brazil |
| andressa.katiski@gmail.com | |
| Phone | 55 41 991922779 |
| Submit Date | 2021-11-12 |
| Num Groups | 7 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | cdf, raw(Waters) |
| Analysis Type Detail | API-MS |
| Release Date | 2023-05-12 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR001264 |
| Project DOI: | doi: 10.21228/M8B39F |
| Project Title: | Metabolomics of the interaction between a consortium of entomopathogenic fungi and their target insect: mechanisms of attack and survival |
| Project Type: | Untargeted Metabolomics |
| Project Summary: | One of the most concerning pests that attack strawberries in Brazil is Duponchelia fovealis, a non-native moth with no registered control methods to date. Our group recently observed that a fungal consortium formed by two strains of Beauveria bassiana increased the mortality of D. fovealis more than inoculation with each strain on its own. However, the molecular interaction between the fungal consortium and the caterpillars is unknown, raising several questions about the enhanced pest control observed. Furthermore, concerns over the emergency of resistance and the selection for resistance to chemical and biological products that are constantly applied in agriculture highlight the need for careful examination of novel pest control methods. Thus, in this work, we sought to pioneer the evaluation of the molecular interaction between a fungal consortium of B. bassiana and D. fovealis caterpillars. We aimed to understand the biocontrol process involved in this interaction and the defense system of the caterpillar. Therefore, seven days after D. fovealis caterpillars were inoculated with the B. bassiana consortium, the dead and surviving caterpillars were analyzed using GC-MS and LC-MS/MS. |
| Institute: | Universidade Federal do Paraná |
| Department: | Patologia Básica |
| Laboratory: | Laboratório de Microbiologia e Biologia Molecular |
| Last Name: | Katiski da Costa Stuart |
| First Name: | Andressa |
| Address: | Av. Cel. Francisco Heráclito dos Santos, 100, Curitiba, Paraná, 81530-000, Brazil |
| Email: | andressa.katiski@gmail.com |
| Phone: | 5541991922779 |
| Project Comments: | Previous studies by our research group showed the increased potential for biocontrol on the insect pest Duponchelia fovealis when two strains of Beauveria bassiana were used together as a fungal consortium. In this work we sought to identify the metabolites involved in the interaction between the consortium formed by different B. bassiana strains and its target insect, D. fovealis. We identify the metabolites using non-targeted metabolomics, applying gas and liquid chromatography coupled to mass spectrometers (GC-MS and LC-MS/MS). These analyses aimed to elucidate the molecular mechanisms involved in the biocontrol effect of the fungal consortium on D. fovealis and to examine possible explanations for the survival of some caterpillars due to a potential resistance mechanism. Dead and surviving caterpillars were analyzed separately. |
| Contributors: | Jason Lee Furuie, Thais Regiani Cataldi, Rodrigo Makowiecky Stuart, Maria Aparecida Cassilha Zawadneak, Carlos Alberto Labate, Ida Chapaval Pimentel |
Subject:
| Subject ID: | SU002071 |
| Subject Type: | Insect |
| Subject Species: | Duponchelia fovealis |
Factors:
Subject type: Insect; Subject species: Duponchelia fovealis (Factor headings shown in green)
| mb_sample_id | local_sample_id | Factor |
|---|---|---|
| SA185990 | CatS_2 | Control |
| SA185991 | CatS_4 | Control |
| SA185992 | CatS_5 | Control |
| SA185993 | CatS_1 | Control |
| SA185994 | CatS_3 | Control |
| SA185995 | CatM_Bov2_1 | Duponchelia fovealis killed by the Bov2 strain of Beauveria bassiana |
| SA185996 | CatM_Bov2_2 | Duponchelia fovealis killed by the Bov2 strain of Beauveria bassiana |
| SA185997 | CatM_Bov2_5 | Duponchelia fovealis killed by the Bov2 strain of Beauveria bassiana |
| SA185998 | CatM_Bov2_3 | Duponchelia fovealis killed by the Bov2 strain of Beauveria bassiana |
| SA185999 | CatM_Bov2_4 | Duponchelia fovealis killed by the Bov2 strain of Beauveria bassiana |
| SA186000 | CatM_Bov3_5 | Duponchelia fovealis killed by the Bov3 strain of Beauveria bassiana |
| SA186001 | CatM_Bov3_4 | Duponchelia fovealis killed by the Bov3 strain of Beauveria bassiana |
| SA186002 | CatM_Bov3_2 | Duponchelia fovealis killed by the Bov3 strain of Beauveria bassiana |
| SA186003 | CatM_Bov3_3 | Duponchelia fovealis killed by the Bov3 strain of Beauveria bassiana |
| SA186004 | CatM_Bov3_1 | Duponchelia fovealis killed by the Bov3 strain of Beauveria bassiana |
| SA186005 | CatM_Cons_3 | Duponchelia fovealis killed by the consortium |
| SA186006 | CatM_Cons_2 | Duponchelia fovealis killed by the consortium |
| SA186007 | CatM_Cons_4 | Duponchelia fovealis killed by the consortium |
| SA186008 | CatM_Cons_5 | Duponchelia fovealis killed by the consortium |
| SA186009 | CatM_Cons_1 | Duponchelia fovealis killed by the consortium |
| SA186010 | CatS_Bov2_1 | Duponchelia fovealis that survived the application of the Bov 2 strain of Beauveria bassiana |
| SA186011 | CatS_Bov2_4 | Duponchelia fovealis that survived the application of the Bov 2 strain of Beauveria bassiana |
| SA186012 | CatS_Bov2_3 | Duponchelia fovealis that survived the application of the Bov 2 strain of Beauveria bassiana |
| SA186013 | CatS_Bov2_2 | Duponchelia fovealis that survived the application of the Bov 2 strain of Beauveria bassiana |
| SA186014 | CatS_Bov2_5 | Duponchelia fovealis that survived the application of the Bov 2 strain of Beauveria bassiana |
| SA186015 | CatS_Bov3_1 | Duponchelia fovealis that survived the application of the Bov 3 strain of Beauveria bassiana |
| SA186016 | CatS_Bov3_3 | Duponchelia fovealis that survived the application of the Bov 3 strain of Beauveria bassiana |
| SA186017 | CatS_Bov3_5 | Duponchelia fovealis that survived the application of the Bov 3 strain of Beauveria bassiana |
| SA186018 | CatS_Bov3_4 | Duponchelia fovealis that survived the application of the Bov 3 strain of Beauveria bassiana |
| SA186019 | CatS_Bov3_2 | Duponchelia fovealis that survived the application of the Bov 3 strain of Beauveria bassiana |
| SA186020 | CatS_Cons_1 | Duponchelia fovealis that survived the application of the consortium |
| SA186021 | CatS_Cons_2 | Duponchelia fovealis that survived the application of the consortium |
| SA186022 | CatS_Cons_3 | Duponchelia fovealis that survived the application of the consortium |
| SA186023 | CatS_Cons_4 | Duponchelia fovealis that survived the application of the consortium |
| SA186024 | CatS_Cons_5 | Duponchelia fovealis that survived the application of the consortium |
| Showing results 1 to 35 of 35 |
Collection:
| Collection ID: | CO002064 |
| Collection Summary: | Groups of 10 third instar caterpillars were placed on strawberry leaves and sprayed with 1 mL of B. bassiana conidia-suspension: 2 x 107 conidia/mL in 0.85% saline solution with Tween 80® adhesive spreader added. Therefore, the treatments applied were: control caterpillars (consisting only of 0.85% saline solution and Tween 80®), caterpillars inoculated with Bov 3 strain of B. bassiana, caterpillars inoculated with Bov 2 strain of B. bassiana and caterpillars inoculated with the Bov3-Bov 2 consortium. On the seventh day, caterpillars that did not respond to the stimulus provided by the touch of a brush were considered dead. All caterpillars were identified as living or dead at the end of the seven days and were subsequently stored in a freezer at -80ºC. |
| Collection Protocol Filename: | Metabolite_Extraction |
| Sample Type: | Insect tissue |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR002083 |
| Treatment Summary: | The treatments applied were: control caterpillars (consisting only of 0.85% saline solution and Tween 80®), caterpillars inoculated with Bov 3 strain of B. bassiana, caterpillars inoculated with Bov 2 strain of B. bassiana and caterpillars inoculated with the Bov3-Bov2 consortium. |
| Treatment Protocol Filename: | Metabolite_Extraction |
| Treatment Dosevolume: | 2 x 10^7 conidia/mL |
Sample Preparation:
| Sampleprep ID: | SP002077 |
| Sampleprep Summary: | After the direct contact bioassay, the caterpillars were arranged by treatment, then macerated in liquid nitrogen (N2). Extraction was performed with 200 mg of the macerate added to a 1 mL microtube (Eppendorf, Germany) previously treated with methanol. Following this, 125 μL of chloroform (CHCl3), 50 μL ultra-pure water (H2O), and 250 μL cold methanol (CH3OH) were added to the macerate. The microtubes were vigorously vortexed and placed in an ultrasonic bath (Odontobrás, Ribeirão-SP) at 20 Hz and approximately 4ºC for 10 minutes. Then, 50 μL of CHCl3 and 50 μL of H2O were added, and the tubes were vortexed again. The samples were centrifuged (Eppendorf, Germany) for 5 minutes at 14000 rpm and 4°C, and the supernatant was filtered on a Whatman® 0.22 µm filter (Merck, Germany) and transferred to a glass vial. The vial was taken to a lyophilizer (Thermo Fischer Scientific, MA) until the samples had completely dried. Finally, the lyophilized samples were resuspended in 200 µL of extraction solution and aliquoted for use in the GC-MS and LC-MS/MS. |
| Sampleprep Protocol Filename: | Metabolite_Extraction |
| Extract Storage: | -80℃ |
Combined analysis:
| Analysis ID | AN003242 | AN003243 | AN003244 | AN003245 | AN003246 | AN003247 |
|---|---|---|---|---|---|---|
| Analysis type | MS | MS | MS | MS | MS | MS |
| Chromatography type | GC | GC | Reversed phase | Reversed phase | Reversed phase | Reversed phase |
| Chromatography system | Agilent 7890A | Agilent 7890A | Waters Acquity UPLC | Waters Acquity UPLC | Waters Acquity UPLC | Waters Acquity UPLC |
| Column | Agilent DB-5 (20m x 0.18mm, 0.18um); Restek RX-T 17 (0.9m x 0.10mm, 0.10um) | Agilent DB-5 (20m x 0.18mm, 0.18um); Restek RX-T 17 (0.9m x 0.10mm, 0.10um) | Waters Acquity UPLC HSS (100 x 2.1mm, 1.7um) | Waters Acquity UPLC HSS (100 x 2.1mm, 1.7um) | Waters Acquity UPLC HSS (100 x 2.1mm, 1.7um) | Waters Acquity UPLC HSS (100 x 2.1mm, 1.7um) |
| MS Type | EI | API | ESI | ESI | ESI | ESI |
| MS instrument type | GC x GC-TOF | GC x GC-TOF | QTOF | QTOF | QTOF | QTOF |
| MS instrument name | Leco Pegasus 4D GCxGC TOF | Leco Pegasus 4D GCxGC TOF | Waters Acquity UPLC | Waters Acquity UPLC | Waters Acquity UPLC | Waters Acquity UPLC |
| Ion Mode | UNSPECIFIED | UNSPECIFIED | NEGATIVE | POSITIVE | NEGATIVE | POSITIVE |
| Units | peak area | Relative intensity | Relative intensity | Relative intensity | Relative intensity |
Chromatography:
| Chromatography ID: | CH002391 |
| Methods Filename: | Metabolomics_Methods |
| Instrument Name: | Agilent 7890A |
| Column Name: | Agilent DB-5 (20m x 0.18mm, 0.18um); Restek RX-T 17 (0.9m x 0.10mm, 0.10um) |
| Column Temperature: | 70 - 320 ºC |
| Flow Rate: | 1 mL.min-1 |
| Injection Temperature: | 280 ºC |
| Sample Injection: | 1 uL |
| Oven Temperature: | 70°C for 2 min, increasing by 15°C·min-1 until it reached 320°C and then held at this temperature for 4 min. |
| Chromatography Type: | GC |
| Chromatography ID: | CH002392 |
| Methods Filename: | Metabolomics_Methods |
| 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. |
| Capillary Voltage: | 3 kV |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS003015 |
| Analysis ID: | AN003242 |
| Instrument Name: | Leco Pegasus 4D GCxGC TOF |
| Instrument Type: | GC x GC-TOF |
| MS Type: | EI |
| MS Comments: | Data from GC-MS was processed using ChromaTOF 4.32 software to conduct baseline correction, deconvolution, retention index (RI), retention time correction (RT), identification, and alignment of peaks. NIST library version 11 was used for the identification of metabolites. Only metabolites with a score of 700 or above were considered. The intensity of each metabolite was normalized by the total ion count (TIC) of each sample. Statistical analyses were performed using the MetaboAnalyst 4.0 online software (available at http://www.metaboanalyst.ca/MetaboAnalyst/) |
| Ion Mode: | UNSPECIFIED |
| Fragmentation Method: | EI |
| Ion Source Temperature: | 250 ºC |
| Ionization Energy: | 70 eV |
| Analysis Protocol File: | metabolomics_methods.pdf |
| MS ID: | MS003016 |
| Analysis ID: | AN003243 |
| Instrument Name: | Leco Pegasus 4D GCxGC TOF |
| Instrument Type: | GC x GC-TOF |
| MS Type: | API |
| MS Comments: | Data from GC-MS was processed using ChromaTOF 4.32 software to conduct baseline correction, deconvolution, retention index (RI), retention time correction (RT), identification, and alignment of peaks. NIST library version 11 was used for the identification of metabolites. Only metabolites with a score of 700 or above were considered. The intensity of each metabolite was normalized by the total ion count (TIC) of each sample. Statistical analyses were performed using the MetaboAnalyst 4.0 online software (available at http://www.metaboanalyst.ca/MetaboAnalyst/) |
| Ion Mode: | UNSPECIFIED |
| Fragmentation Method: | EI |
| Ion Source Temperature: | 250 ºC |
| Ionization Energy: | 70 eV |
| Analysis Protocol File: | metabolomics_methods.pdf |
| MS ID: | MS003017 |
| Analysis ID: | AN003244 |
| Instrument Name: | Waters Acquity UPLC |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | Generated data were pre-processed using MassLynx 4.1 software (Waters Corporation, MA, USA) and then analyzed using MetaboAnalyst 4.0 online software. Fragmentation was performed under the same conditions as the ionization source, using collision energies between 15 and 50 eV. The search for metabolites was performed in the Human Metabolome Database (HMDB) using a mass tolerance of up to 0.1 Da and considering the adduct of [M-H]-. The structures of the molecules were imported and underwent in silico fragmentation using ACD/MS Structure ID software suite (ACD/labs, Toronto, Canada). The fragmentation profile of each molecule proposed by the program was then compared to the fragments generated by MS/MS to accept or reject the identification of metabolites according to similarity. |
| Ion Mode: | NEGATIVE |
| Capillary Voltage: | 3 kV |
| Dry Gas Flow: | 50 L/hr |
| Source Temperature: | 150 ºC |
| Desolvation Gas Flow: | 550 L/hr. |
| Analysis Protocol File: | metabolomics_methods.pdf |
| MS ID: | MS003018 |
| Analysis ID: | AN003245 |
| Instrument Name: | Waters Acquity UPLC |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | Generated data were pre-processed using MassLynx 4.1 software (Waters Corporation, MA, USA) and then analyzed using MetaboAnalyst 4.0 online software. Fragmentation was performed under the same conditions as the ionization source, using collision energies between 15 and 50 eV. The search for metabolites was performed in the Human Metabolome Database (HMDB) using a mass tolerance of up to 0.1 Da and considering the adduct of [M-H]-. The structures of the molecules were imported and underwent in silico fragmentation using ACD/MS Structure ID software suite (ACD/labs, Toronto, Canada). The fragmentation profile of each molecule proposed by the program was then compared to the fragments generated by MS/MS to accept or reject the identification of metabolites according to similarity. |
| Ion Mode: | POSITIVE |
| Capillary Voltage: | 3 kV |
| Dry Gas Flow: | 50 L/hr |
| Source Temperature: | 150 ºC |
| Desolvation Gas Flow: | 550 L/hr |
| Analysis Protocol File: | metabolomics_methods.pdf |
| MS ID: | MS003019 |
| Analysis ID: | AN003246 |
| Instrument Name: | Waters Acquity UPLC |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | Generated data were pre-processed using MassLynx 4.1 software (Waters Corporation, MA, USA) and then analyzed using MetaboAnalyst 4.0 online software. Fragmentation was performed under the same conditions as the ionization source, using collision energies between 15 and 50 eV. The search for metabolites was performed in the Human Metabolome Database (HMDB) using a mass tolerance of up to 0.1 Da and considering the adduct of [M-H]-. The structures of the molecules were imported and underwent in silico fragmentation using ACD/MS Structure ID software suite (ACD/labs, Toronto, Canada). The fragmentation profile of each molecule proposed by the program was then compared to the fragments generated by MS/MS to accept or reject the identification of metabolites according to similarity. |
| Ion Mode: | NEGATIVE |
| Capillary Voltage: | 3 kV |
| Dry Gas Flow: | 50 L/hr |
| Source Temperature: | 150 ºC |
| Desolvation Gas Flow: | 550 L/hr |
| Analysis Protocol File: | metabolomics_methods.pdf |
| MS ID: | MS003020 |
| Analysis ID: | AN003247 |
| Instrument Name: | Waters Acquity UPLC |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | Generated data were pre-processed using MassLynx 4.1 software (Waters Corporation, MA, USA) and then analyzed using MetaboAnalyst 4.0 online software. Fragmentation was performed under the same conditions as the ionization source, using collision energies between 15 and 50 eV. The search for metabolites was performed in the Human Metabolome Database (HMDB) using a mass tolerance of up to 0.1 Da and considering the adduct of [M-H]-. The structures of the molecules were imported and underwent in silico fragmentation using ACD/MS Structure ID software suite (ACD/labs, Toronto, Canada). The fragmentation profile of each molecule proposed by the program was then compared to the fragments generated by MS/MS to accept or reject the identification of metabolites according to similarity. |
| Ion Mode: | POSITIVE |
| Capillary Voltage: | 3 kV |
| Dry Gas Flow: | 50 L/hr |
| Source Temperature: | 150 ºC |
| Desolvation Gas Flow: | 550 L/hr |
| Analysis Protocol File: | metabolomics_methods.pdf |
