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MB Sample ID: SA078174
| Local Sample ID: | C9 |
| Subject ID: | SU001195 |
| Subject Type: | Plant |
| Subject Species: | Brassica nigra |
| Taxonomy ID: | 3710 |
| Age Or Age Range: | 4 weeks old |
| Height Or Height Range: | 15-20 cm |
| Species Group: | Plants |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001195 |
| Subject Type: | Plant |
| Subject Species: | Brassica nigra |
| Taxonomy ID: | 3710 |
| Age Or Age Range: | 4 weeks old |
| Height Or Height Range: | 15-20 cm |
| Species Group: | Plants |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| C9 | SA078174 | FL011912 | control | Treatments |
Collection:
| Collection ID: | CO001189 |
| Collection Summary: | Leaf samples were harvested by cutting the petiole with scissors, wrapping the leaves in aluminium foil labelled with the plant's ID and leaf position, and immediately flash-freezing them in liquid nitrogen. Samples were then stored at -80°C until being further prepared for metabolic analysis. |
| Sample Type: | Plant |
| Collection Location: | greenhouse |
| Collection Duration: | 1 hour |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR001210 |
| Treatment Summary: | Two types of treatment were used separately and in sequence: simulated herbivory with use of the phytohormone MeJA (M); real herbivory by P. brassicae caterpillars (P); and combined treatment (MP) where caterpillars were introduced to feed on (M)-plants for five days, after three days of MeJA pre-treatment. MeJA (Sigma-Aldrich, CAS Number 39924-52-2) was applied to the whole plant by spraying 5 mL (1mM, diluted in MilliQ water). First instar caterpillars (15 individuals per plant) were placed on the first fully expanded leaf (L4) of experimental plant and left to feed freely for five days. |
| Plant Light Period: | 16:8 hours light: dark cycle |
| Plant Humidity: | 50-70% RH |
| Plant Temp: | 20-22 °C |
| Plant Growth Stage: | vegetative (4 weeks) |
Sample Preparation:
| Sampleprep ID: | SP001203 |
| Sampleprep Summary: | Each sample (10-12 mg) was extracted in 1 ml of cold chloroform:methanol:H2O (20:60:20 v/v), including 7.5 ng/µl of the isotope-labelled internal standard, SA-D4. Extracts were agitated with a 3 mm tungsten carbide bead for 3 min and centrifuged at 20,800 × g for 10 min at 4°C. The supernatant (200 μl) was dried and processed for GC/MS analyses |
| Processing Storage Conditions: | On ice |
| Extract Storage: | -80℃ |
| Sample Derivatization: | Dry extracts were derivatized with methoxyamine and MSTFA |
Combined analysis:
| Analysis ID | AN001859 |
|---|---|
| Analysis type | MS |
| Chromatography type | GC |
| Chromatography system | Agilent 6890 |
| Column | DB5-MS |
| MS Type | EI |
| MS instrument type | GC-TOF |
| MS instrument name | Agilent 6890 |
| Ion Mode | POSITIVE |
| Units | Normalized peak area |
Chromatography:
| Chromatography ID: | CH001346 |
| Chromatography Summary: | Samples were analysed by GC-TOF-MS (on an Agilent 6890 gas chromatograph equipped with a 10 m x 0.18 mm silica capillary column with a 0.18 µm DB 5-MS UI stationary phase, J&W Scientific; connected to a LECO Pegasus III TOF-MS) operated using the LECO ChromaTOF® software package (Leco Corp., St Joseph, MI, USA). Retention indices (RIs) were calculated relative to those of a C8-C40 alkane series. Deconvolution, feature extraction and peak integration were performed using a Matlab® script developed in-house. Identification was achieved by comparing compounds' RIs and mass spectra to the SMC in-house mass spectra library and to the public Golm Metabolome Database developed by the Max Planck Institute (GMD-MPI; http://gmd.mpimp-golm.mpg.de). Samples were normalized with respect to the peak areas of the internal standards (methyl stearate and SA-D4). |
| Instrument Name: | Agilent 6890 |
| Column Name: | DB5-MS |
| Flow Rate: | 1mL/min |
| Injection Temperature: | 270°C |
| Internal Standard: | salicylic acid |
| Internal Standard Mt: | stearic acid |
| Capillary Voltage: | 70-eV |
| Oven Temperature: | 70°C to 320°C (40 °C /min) |
| Running Voltage: | detector voltage 1,700V. |
| Transferline Temperature: | 250°C |
| Chromatography Type: | GC |
MS:
| MS ID: | MS001719 |
| Analysis ID: | AN001859 |
| Instrument Name: | Agilent 6890 |
| Instrument Type: | GC-TOF |
| MS Type: | EI |
| MS Comments: | Samples were analysed by GC-TOF-MS (on an Agilent 6890 gas chromatograph equipped with a 10 m x 0.18 mm silica capillary column with a 0.18 µm DB 5-MS UI stationary phase, J&W Scientific; connected to a LECO Pegasus III TOF-MS) operated using the LECO ChromaTOF® software package (Leco Corp., St Joseph, MI, USA). Retention indices (RIs) were calculated relative to those of a C8-C40 alkane series. Deconvolution, feature extraction and peak integration were performed using a Matlab® script developed in-house. Identification was achieved by comparing compounds' RIs and mass spectra to the SMC in-house mass spectra library and to the public Golm Metabolome Database developed by the Max Planck Institute (GMD-MPI; http://gmd.mpimp-golm.mpg.de). Samples were normalized with respect to the peak areas of the internal standards (methyl stearate and SA-D4). |
| Ion Mode: | POSITIVE |
