Study Data
| Project uploaded by: | Yashwant |
| Project ID: | IMP_100050 |
| Title: | Metabolic and Lipidomic Trade-offs in Helicoverpa armigera: Dynamics Under Plant Protease Inhibitor-Induced Stress |
| Project Description: | Plant protease inhibitors retard the growth and development of insects by inhibiting their digestive proteases. In response, insects try to adapt to these plant defensive molecules by modulating their protease expression. However, their survival mechanisms might not be limited only to digestive plasticity. To explore this, we performed a comprehensive lipidomics and metabolomics analysis in Helicoverpa armigera fed with a recombinant Capsicum annuum protease inhibitor (rCanPI-7) having unique four inhibitory repeat domains with potent activity against insect trypsins and chymotrypsins. These results revealed that H. armigera employs a dynamic and multifaceted physiological response to dietary stress induced by rCanPI. Upon ingestion of rCanPI-7, down regulation of glycolysis and TCA cycle indicated a decrease in primary energy metabolism while oxidative stress was evident from the depletion of reduced glutathione, peroxidation of membrane lipids, and accumulation of ceramides which are the hallmarks of mitochondrial dysfunction. Investigation of the dynamics in the turnover of different molecules hints that H. armigera activated multiple compensatory strategies such as mobilizing triglycerides and amino acid catabolism as an alternative source of energy, upregulation of antioxidants, membrane remodeling, activation of apoptosis, and shifts in neuromodulatory metabolites linked to cognitive adaptation. Collectively, these findings point to a tightly regulated physiological tug-of-war in H. armigera, where the damaging impact of rCanPI-induced oxidative and nutritional stress is counteracted by a suite of compensatory metabolic, structural, and neuromodulatory adjustments. To our knowledge, this is the first report of lipidomic profiling in H. armigera, providing novel insights into its biochemical resilience and identifying potential metabolic vulnerabilities for enhancing biopesticide strategies. |
| Research Area: | Biological Sciences |
| Funding Source: | Translational Research Program (TRP) (No. BT/PR30159/MED/15/188/2018) of Department of Biotechnology (DBT), Govt. of India. |
| Project Contributors: | Yashwant Kumar |
| Sr.No | Sample ID | Sample Name | Organism | Source | Sample Preparation Protocol | Sample Type | Experimental Condition | Time of treatment | Variant/Variety | Gender | Age | Replicates | Storage Conditions | Extraction Protocol | Number of files per sample |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| 11 | IMSM_102228 | LI_2 | Helicoverpa armigera | 29058 | Insects | Experiment design and feeding assays were performed as per our previous study (Lomate et al., 2018). In brief, H. armigera larvae were maintained at optimal growth conditions in the laboratory with 27 ± 2°C, 60 ± 5% relative humidity and a photoperiod of 14 h light and 10 h dark. An artificial diet (AD) was prepared as per (Mahajan et al., 2013), and the PI diet was prepared by adding 150 μg of recombinant Capsicum annum protease inhibitor (rCanPI-7) to the artificial diet. Neonates were fed on artificial diet for 2 days, and then first instar larvae were transferred to the control artificial diet (AD-fed) and rCanPI-7 incorporated artificial diet (CanPI-fed) for 48 hours. Whole larvae were harvested at 0.5, 2, 6, 12, 24 and 48 h, each set containing 100 larvae. Pooled samples of 0.5, 2, and 6 h (early response), 12 and 24 h (mid response), and 48 h (late response) were studied using lipidomic and metabolomic studies. At each stage of bioassay, the harvested samples were snap frozen in liquid nitrogen and stored at -80°C until further use. Three biological replicates were used for both lipidomic and metabolomic study. | Late Fed | Fed | 48 hrs | NA | NA | NA | 3 biological replicates | -80°C |
Metabolites were extracted by adding 500 µL of 80% chilled methanol (MS-grade, Waters) to 25 mg of frozen and crushed tissue. The suspension was vortexed for 1 min and frozen at -80°C for 10 min. The freeze-thaw cycle was repeated twice, followed by centrifugation at 15,000g for 10 min at 4°C. The supernatant was collected in a separate tube, and 100 μL was dried using a speed vacuum at room temperature for 20 to 25 min. Samples were stored at -80°C till further analysis. For sample injection, each sample was re-suspended in 25 μL of methanol-water mixture (3:17, methanol: water), vortexed briefly for 30 s, and centrifuged at 14,000 rpm for 10 min at 4°C. |
4 |
| 12 | IMSM_102229 | LI_3 | Helicoverpa armigera | 29058 | Insects | Experiment design and feeding assays were performed as per our previous study (Lomate et al., 2018). In brief, H. armigera larvae were maintained at optimal growth conditions in the laboratory with 27 ± 2°C, 60 ± 5% relative humidity and a photoperiod of 14 h light and 10 h dark. An artificial diet (AD) was prepared as per (Mahajan et al., 2013), and the PI diet was prepared by adding 150 μg of recombinant Capsicum annum protease inhibitor (rCanPI-7) to the artificial diet. Neonates were fed on artificial diet for 2 days, and then first instar larvae were transferred to the control artificial diet (AD-fed) and rCanPI-7 incorporated artificial diet (CanPI-fed) for 48 hours. Whole larvae were harvested at 0.5, 2, 6, 12, 24 and 48 h, each set containing 100 larvae. Pooled samples of 0.5, 2, and 6 h (early response), 12 and 24 h (mid response), and 48 h (late response) were studied using lipidomic and metabolomic studies. At each stage of bioassay, the harvested samples were snap frozen in liquid nitrogen and stored at -80°C until further use. Three biological replicates were used for both lipidomic and metabolomic study. | Late Fed | Fed | 48 hrs | NA | NA | NA | 3 biological replicates | -80°C |
Metabolites were extracted by adding 500 µL of 80% chilled methanol (MS-grade, Waters) to 25 mg of frozen and crushed tissue. The suspension was vortexed for 1 min and frozen at -80°C for 10 min. The freeze-thaw cycle was repeated twice, followed by centrifugation at 15,000g for 10 min at 4°C. The supernatant was collected in a separate tube, and 100 μL was dried using a speed vacuum at room temperature for 20 to 25 min. Samples were stored at -80°C till further analysis. For sample injection, each sample was re-suspended in 25 μL of methanol-water mixture (3:17, methanol: water), vortexed briefly for 30 s, and centrifuged at 14,000 rpm for 10 min at 4°C. |
4 |
| 13 | IMSM_102230 | MC_1 | Helicoverpa armigera | 29058 | Insects | Experiment design and feeding assays were performed as per our previous study (Lomate et al., 2018). In brief, H. armigera larvae were maintained at optimal growth conditions in the laboratory with 27 ± 2°C, 60 ± 5% relative humidity and a photoperiod of 14 h light and 10 h dark. An artificial diet (AD) was prepared as per (Mahajan et al., 2013), and the PI diet was prepared by adding 150 μg of recombinant Capsicum annum protease inhibitor (rCanPI-7) to the artificial diet. Neonates were fed on artificial diet for 2 days, and then first instar larvae were transferred to the control artificial diet (AD-fed) and rCanPI-7 incorporated artificial diet (CanPI-fed) for 48 hours. Whole larvae were harvested at 0.5, 2, 6, 12, 24 and 48 h, each set containing 100 larvae. Pooled samples of 0.5, 2, and 6 h (early response), 12 and 24 h (mid response), and 48 h (late response) were studied using lipidomic and metabolomic studies. At each stage of bioassay, the harvested samples were snap frozen in liquid nitrogen and stored at -80°C until further use. Three biological replicates were used for both lipidomic and metabolomic study. | Mid Control | Control | 48 hrs | NA | NA | NA | 3 biological replicates | -80°C |
Metabolites were extracted by adding 500 µL of 80% chilled methanol (MS-grade, Waters) to 25 mg of frozen and crushed tissue. The suspension was vortexed for 1 min and frozen at -80°C for 10 min. The freeze-thaw cycle was repeated twice, followed by centrifugation at 15,000g for 10 min at 4°C. The supernatant was collected in a separate tube, and 100 μL was dried using a speed vacuum at room temperature for 20 to 25 min. Samples were stored at -80°C till further analysis. For sample injection, each sample was re-suspended in 25 μL of methanol-water mixture (3:17, methanol: water), vortexed briefly for 30 s, and centrifuged at 14,000 rpm for 10 min at 4°C. |
4 |
| 14 | IMSM_102231 | MC_2 | Helicoverpa armigera | 29058 | Insects | Experiment design and feeding assays were performed as per our previous study (Lomate et al., 2018). In brief, H. armigera larvae were maintained at optimal growth conditions in the laboratory with 27 ± 2°C, 60 ± 5% relative humidity and a photoperiod of 14 h light and 10 h dark. An artificial diet (AD) was prepared as per (Mahajan et al., 2013), and the PI diet was prepared by adding 150 μg of recombinant Capsicum annum protease inhibitor (rCanPI-7) to the artificial diet. Neonates were fed on artificial diet for 2 days, and then first instar larvae were transferred to the control artificial diet (AD-fed) and rCanPI-7 incorporated artificial diet (CanPI-fed) for 48 hours. Whole larvae were harvested at 0.5, 2, 6, 12, 24 and 48 h, each set containing 100 larvae. Pooled samples of 0.5, 2, and 6 h (early response), 12 and 24 h (mid response), and 48 h (late response) were studied using lipidomic and metabolomic studies. At each stage of bioassay, the harvested samples were snap frozen in liquid nitrogen and stored at -80°C until further use. Three biological replicates were used for both lipidomic and metabolomic study. | Mid Control | Control | 48 hrs | NA | NA | NA | 3 biological replicates | -80°C |
Metabolites were extracted by adding 500 µL of 80% chilled methanol (MS-grade, Waters) to 25 mg of frozen and crushed tissue. The suspension was vortexed for 1 min and frozen at -80°C for 10 min. The freeze-thaw cycle was repeated twice, followed by centrifugation at 15,000g for 10 min at 4°C. The supernatant was collected in a separate tube, and 100 μL was dried using a speed vacuum at room temperature for 20 to 25 min. Samples were stored at -80°C till further analysis. For sample injection, each sample was re-suspended in 25 μL of methanol-water mixture (3:17, methanol: water), vortexed briefly for 30 s, and centrifuged at 14,000 rpm for 10 min at 4°C. |
4 |
| 15 | IMSM_102232 | MC_3 | Helicoverpa armigera | 29058 | Insects | Experiment design and feeding assays were performed as per our previous study (Lomate et al., 2018). In brief, H. armigera larvae were maintained at optimal growth conditions in the laboratory with 27 ± 2°C, 60 ± 5% relative humidity and a photoperiod of 14 h light and 10 h dark. An artificial diet (AD) was prepared as per (Mahajan et al., 2013), and the PI diet was prepared by adding 150 μg of recombinant Capsicum annum protease inhibitor (rCanPI-7) to the artificial diet. Neonates were fed on artificial diet for 2 days, and then first instar larvae were transferred to the control artificial diet (AD-fed) and rCanPI-7 incorporated artificial diet (CanPI-fed) for 48 hours. Whole larvae were harvested at 0.5, 2, 6, 12, 24 and 48 h, each set containing 100 larvae. Pooled samples of 0.5, 2, and 6 h (early response), 12 and 24 h (mid response), and 48 h (late response) were studied using lipidomic and metabolomic studies. At each stage of bioassay, the harvested samples were snap frozen in liquid nitrogen and stored at -80°C until further use. Three biological replicates were used for both lipidomic and metabolomic study. | Mid Control | Control | 48 hrs | NA | NA | NA | 3 biological replicates | -80°C |
Metabolites were extracted by adding 500 µL of 80% chilled methanol (MS-grade, Waters) to 25 mg of frozen and crushed tissue. The suspension was vortexed for 1 min and frozen at -80°C for 10 min. The freeze-thaw cycle was repeated twice, followed by centrifugation at 15,000g for 10 min at 4°C. The supernatant was collected in a separate tube, and 100 μL was dried using a speed vacuum at room temperature for 20 to 25 min. Samples were stored at -80°C till further analysis. For sample injection, each sample was re-suspended in 25 μL of methanol-water mixture (3:17, methanol: water), vortexed briefly for 30 s, and centrifuged at 14,000 rpm for 10 min at 4°C. |
4 |
| 16 | IMSM_102233 | MI_1 | Helicoverpa armigera | 29058 | Insects | Experiment design and feeding assays were performed as per our previous study (Lomate et al., 2018). In brief, H. armigera larvae were maintained at optimal growth conditions in the laboratory with 27 ± 2°C, 60 ± 5% relative humidity and a photoperiod of 14 h light and 10 h dark. An artificial diet (AD) was prepared as per (Mahajan et al., 2013), and the PI diet was prepared by adding 150 μg of recombinant Capsicum annum protease inhibitor (rCanPI-7) to the artificial diet. Neonates were fed on artificial diet for 2 days, and then first instar larvae were transferred to the control artificial diet (AD-fed) and rCanPI-7 incorporated artificial diet (CanPI-fed) for 48 hours. Whole larvae were harvested at 0.5, 2, 6, 12, 24 and 48 h, each set containing 100 larvae. Pooled samples of 0.5, 2, and 6 h (early response), 12 and 24 h (mid response), and 48 h (late response) were studied using lipidomic and metabolomic studies. At each stage of bioassay, the harvested samples were snap frozen in liquid nitrogen and stored at -80°C until further use. Three biological replicates were used for both lipidomic and metabolomic study. | Mid Fed | Fed | 48 hrs | NA | NA | NA | 3 biological replicates | -80°C |
Metabolites were extracted by adding 500 µL of 80% chilled methanol (MS-grade, Waters) to 25 mg of frozen and crushed tissue. The suspension was vortexed for 1 min and frozen at -80°C for 10 min. The freeze-thaw cycle was repeated twice, followed by centrifugation at 15,000g for 10 min at 4°C. The supernatant was collected in a separate tube, and 100 μL was dried using a speed vacuum at room temperature for 20 to 25 min. Samples were stored at -80°C till further analysis. For sample injection, each sample was re-suspended in 25 μL of methanol-water mixture (3:17, methanol: water), vortexed briefly for 30 s, and centrifuged at 14,000 rpm for 10 min at 4°C. |
4 |
| 17 | IMSM_102234 | MI_2 | Helicoverpa armigera | 29058 | Insects | Experiment design and feeding assays were performed as per our previous study (Lomate et al., 2018). In brief, H. armigera larvae were maintained at optimal growth conditions in the laboratory with 27 ± 2°C, 60 ± 5% relative humidity and a photoperiod of 14 h light and 10 h dark. An artificial diet (AD) was prepared as per (Mahajan et al., 2013), and the PI diet was prepared by adding 150 μg of recombinant Capsicum annum protease inhibitor (rCanPI-7) to the artificial diet. Neonates were fed on artificial diet for 2 days, and then first instar larvae were transferred to the control artificial diet (AD-fed) and rCanPI-7 incorporated artificial diet (CanPI-fed) for 48 hours. Whole larvae were harvested at 0.5, 2, 6, 12, 24 and 48 h, each set containing 100 larvae. Pooled samples of 0.5, 2, and 6 h (early response), 12 and 24 h (mid response), and 48 h (late response) were studied using lipidomic and metabolomic studies. At each stage of bioassay, the harvested samples were snap frozen in liquid nitrogen and stored at -80°C until further use. Three biological replicates were used for both lipidomic and metabolomic study. | Mid Fed | Fed | 48 hrs | NA | NA | NA | 3 biological replicates | -80°C |
Metabolites were extracted by adding 500 µL of 80% chilled methanol (MS-grade, Waters) to 25 mg of frozen and crushed tissue. The suspension was vortexed for 1 min and frozen at -80°C for 10 min. The freeze-thaw cycle was repeated twice, followed by centrifugation at 15,000g for 10 min at 4°C. The supernatant was collected in a separate tube, and 100 μL was dried using a speed vacuum at room temperature for 20 to 25 min. Samples were stored at -80°C till further analysis. For sample injection, each sample was re-suspended in 25 μL of methanol-water mixture (3:17, methanol: water), vortexed briefly for 30 s, and centrifuged at 14,000 rpm for 10 min at 4°C. |
4 |
| 18 | IMSM_102235 | MI_3 | Helicoverpa armigera | 29058 | Insects | Experiment design and feeding assays were performed as per our previous study (Lomate et al., 2018). In brief, H. armigera larvae were maintained at optimal growth conditions in the laboratory with 27 ± 2°C, 60 ± 5% relative humidity and a photoperiod of 14 h light and 10 h dark. An artificial diet (AD) was prepared as per (Mahajan et al., 2013), and the PI diet was prepared by adding 150 μg of recombinant Capsicum annum protease inhibitor (rCanPI-7) to the artificial diet. Neonates were fed on artificial diet for 2 days, and then first instar larvae were transferred to the control artificial diet (AD-fed) and rCanPI-7 incorporated artificial diet (CanPI-fed) for 48 hours. Whole larvae were harvested at 0.5, 2, 6, 12, 24 and 48 h, each set containing 100 larvae. Pooled samples of 0.5, 2, and 6 h (early response), 12 and 24 h (mid response), and 48 h (late response) were studied using lipidomic and metabolomic studies. At each stage of bioassay, the harvested samples were snap frozen in liquid nitrogen and stored at -80°C until further use. Three biological replicates were used for both lipidomic and metabolomic study. | Mid Fed | Fed | 48 hrs | NA | NA | NA | 3 biological replicates | -80°C |
Metabolites were extracted by adding 500 µL of 80% chilled methanol (MS-grade, Waters) to 25 mg of frozen and crushed tissue. The suspension was vortexed for 1 min and frozen at -80°C for 10 min. The freeze-thaw cycle was repeated twice, followed by centrifugation at 15,000g for 10 min at 4°C. The supernatant was collected in a separate tube, and 100 μL was dried using a speed vacuum at room temperature for 20 to 25 min. Samples were stored at -80°C till further analysis. For sample injection, each sample was re-suspended in 25 μL of methanol-water mixture (3:17, methanol: water), vortexed briefly for 30 s, and centrifuged at 14,000 rpm for 10 min at 4°C. |
4 |
| Sr.No | MS Exp ID | Sample Name/ID | Mass Spectrometer Type | MS Instrument Name | MS Instrument type | MS Ionization Method | Ion Mode/Scan Polarity | Data Transformation (Software/s Used) |
|---|---|---|---|---|---|---|---|---|
| 21 | IME_101556 | EI_3 / IMSM_102223 | LCMS (Liquid Chromatography- Mass Spectrometry) | Thermo Fusion Tribrid Orbitrap | Orbitrap | Electrospray Ionization - ESI | Negative | NA |
| 22 | IME_101557 | EI_3 / IMSM_102223 | LCMS (Liquid Chromatography- Mass Spectrometry) | Thermo Fusion Tribrid Orbitrap | Orbitrap | Electrospray Ionization - ESI | Positive | NA |
| 23 | IME_101558 | EI_3 / IMSM_102223 | LCMS (Liquid Chromatography- Mass Spectrometry) | Thermo Fusion Tribrid Orbitrap | Orbitrap | Electrospray Ionization - ESI | Negative | NA |
| 24 | IME_101559 | EI_3 / IMSM_102223 | LCMS (Liquid Chromatography- Mass Spectrometry) | Thermo Fusion Tribrid Orbitrap | Orbitrap | Electrospray Ionization - ESI | Positive | NA |
| 25 | IME_101560 | LC_1 / IMSM_102224 | LCMS (Liquid Chromatography- Mass Spectrometry) | Thermo Fusion Tribrid Orbitrap | Orbitrap | Electrospray Ionization - ESI | Negative | NA |
| 26 | IME_101561 | LC_1 / IMSM_102224 | LCMS (Liquid Chromatography- Mass Spectrometry) | Thermo Fusion Tribrid Orbitrap | Orbitrap | Electrospray Ionization - ESI | Positive | NA |
| 27 | IME_101562 | LC_1 / IMSM_102224 | LCMS (Liquid Chromatography- Mass Spectrometry) | Thermo Fusion Tribrid Orbitrap | Orbitrap | Electrospray Ionization - ESI | Negative | NA |
| 28 | IME_101563 | LC_1 / IMSM_102224 | LCMS (Liquid Chromatography- Mass Spectrometry) | Thermo Fusion Tribrid Orbitrap | Orbitrap | Electrospray Ionization - ESI | Positive | NA |
| 29 | IME_101564 | LC_2 / IMSM_102225 | LCMS (Liquid Chromatography- Mass Spectrometry) | Thermo Fusion Tribrid Orbitrap | Orbitrap | Electrospray Ionization - ESI | Negative | NA |
| 30 | IME_101565 | LC_2 / IMSM_102225 | LCMS (Liquid Chromatography- Mass Spectrometry) | Thermo Fusion Tribrid Orbitrap | Orbitrap | Electrospray Ionization - ESI | Positive | NA |
| Sr.No | First name | Last name | Organization | Designation | |
|---|---|---|---|---|---|
| 1 | Yashwant | Kumar | y.kumar@thsti.res.in | Translational Health Science And Technology Institute (THSTI) | principal_investigator |
| Sr.No | ftprun ID | MS Exp ID | MS Data Files |
|---|---|---|---|
| 51 | IMR_102190 | IME_101586 | HILIC_NEG_MC_1.mzXML |
| 52 | IMR_102191 | IME_101587 | HILIC_POS_MC_1.mzXML |
| 53 | IMR_102192 | IME_101588 | RP_NEG_MC_2.mzXML |
| 54 | IMR_102193 | IME_101589 | RP_POS_MC_2.mzXML |
| 55 | IMR_102194 | IME_101590 | HILIC_NEG_MC_2.mzXML |
| 56 | IMR_102195 | IME_101591 | HILIC_POS_MC_2.mzXML |
| 57 | IMR_102196 | IME_101592 | RP_NEG_MC_3.mzXML |
| 58 | IMR_102197 | IME_101593 | RP_POS_MC_3.mzXML |
| 59 | IMR_102198 | IME_101594 | HILIC_NEG_MC_3.mzXML |
| 60 | IMR_102199 | IME_101595 | HILIC_POS_MC_3.mzXML |